Mutagenesis where the mutation is caused by the introduction of foreign DNA sequences into a gene or extragenic sequence. This may occur spontaneously in vivo or be experimentally induced in vivo or in vitro. Proviral DNA insertions into or adjacent to a cellular proto-oncogene can interrupt GENETIC TRANSLATION of the coding sequences or interfere with recognition of regulatory elements and cause unregulated expression of the proto-oncogene resulting in tumor formation.
Molecular Sequence Data
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
Amino Acid Sequence
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
Amino Acid Substitution
The naturally occurring or experimentally induced replacement of one or more AMINO ACIDS in a protein with another. If a functionally equivalent amino acid is substituted, the protein may retain wild-type activity. Substitution may also diminish, enhance, or eliminate protein function. Experimentally induced substitution is often used to study enzyme activities and binding site properties.
Sequence Homology, Amino Acid
DNA Transposable Elements
Discrete segments of DNA which can excise and reintegrate to another site in the genome. Most are inactive, i.e., have not been found to exist outside the integrated state. DNA transposable elements include bacterial IS (insertion sequence) elements, Tn elements, the maize controlling elements Ac and Ds, Drosophila P, gypsy, and pogo elements, the human Tigger elements and the Tc and mariner elements which are found throughout the animal kingdom.
Protein Structure, Tertiary
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
Protein Structure, Secondary
The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.
SOS Response (Genetics)
An error-prone mechanism or set of functions for repairing damaged microbial DNA. SOS functions (a concept reputedly derived from the SOS of the international distress signal) are involved in DNA repair and mutagenesis, in cell division inhibition, in recovery of normal physiological conditions after DNA repair, and possibly in cell death when DNA damage is extensive.
Genetic Complementation Test
Recombinant Fusion Proteins
Procedures by which protein structure and function are changed or created in vitro by altering existing or synthesizing new structural genes that direct the synthesis of proteins with sought-after properties. Such procedures may include the design of MOLECULAR MODELS of proteins using COMPUTER GRAPHICS or other molecular modeling techniques; site-specific mutagenesis (MUTAGENESIS, SITE-SPECIFIC) of existing genes; and DIRECTED MOLECULAR EVOLUTION techniques to create new genes.
Directed Molecular Evolution
The techniques used to produce molecules exhibiting properties that conform to the demands of the experimenter. These techniques combine methods of generating structural changes with methods of selection. They are also used to examine proposed mechanisms of evolution under in vitro selection conditions.
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
Amino Acid Motifs
Promoter Regions, Genetic
That portion of the electromagnetic spectrum immediately below the visible range and extending into the x-ray frequencies. The longer wavelengths (near-UV or biotic or vital rays) are necessary for the endogenous synthesis of vitamin D and are also called antirachitic rays; the shorter, ionizing wavelengths (far-UV or abiotic or extravital rays) are viricidal, bactericidal, mutagenic, and carcinogenic and are used as disinfectants.
Gene Expression Regulation, Bacterial
DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition.
CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)
Polymerase Chain Reaction
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
The reconstruction of a continuous two-stranded DNA molecule without mismatch from a molecule which contained damaged regions. The major repair mechanisms are excision repair, in which defective regions in one strand are excised and resynthesized using the complementary base pairing information in the intact strand; photoreactivation repair, in which the lethal and mutagenic effects of ultraviolet light are eliminated; and post-replication repair, in which the primary lesions are not repaired, but the gaps in one daughter duplex are filled in by incorporation of portions of the other (undamaged) daughter duplex. Excision repair and post-replication repair are sometimes referred to as "dark repair" because they do not require light.
DNA-Directed DNA Polymerase
DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.
Sequence Analysis, DNA
A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed)
An essential amino acid that is necessary for normal growth in infants and for NITROGEN balance in adults. It is a precursor of INDOLE ALKALOIDS in plants. It is a precursor of SEROTONIN (hence its use as an antidepressant and sleep aid). It can be a precursor to NIACIN, albeit inefficiently, in mammals.
A set of three nucleotides in a protein coding sequence that specifies individual amino acids or a termination signal (CODON, TERMINATOR). Most codons are universal, but some organisms do not produce the transfer RNAs (RNA, TRANSFER) complementary to all codons. These codons are referred to as unassigned codons (CODONS, NONSENSE).
Electrophoresis, Polyacrylamide Gel
Injuries to DNA that introduce deviations from its normal, intact structure and which may, if left unrepaired, result in a MUTATION or a block of DNA REPLICATION. These deviations may be caused by physical or chemical agents and occur by natural or unnatural, introduced circumstances. They include the introduction of illegitimate bases during replication or by deamination or other modification of bases; the loss of a base from the DNA backbone leaving an abasic site; single-strand breaks; double strand breaks; and intrastrand (PYRIMIDINE DIMERS) or interstrand crosslinking. Damage can often be repaired (DNA REPAIR). If the damage is extensive, it can induce APOPTOSIS.
A theoretical representative nucleotide or amino acid sequence in which each nucleotide or amino acid is the one which occurs most frequently at that site in the different sequences which occur in nature. The phrase also refers to an actual sequence which approximates the theoretical consensus. A known CONSERVED SEQUENCE set is represented by a consensus sequence. Commonly observed supersecondary protein structures (AMINO ACID MOTIFS) are often formed by conserved sequences.
Rec A Recombinases
A family of recombinases initially identified in BACTERIA. They catalyze the ATP-driven exchange of DNA strands in GENETIC RECOMBINATION. The product of the reaction consists of a duplex and a displaced single-stranded loop, which has the shape of the letter D and is therefore called a D-loop structure.
Protein Processing, Post-Translational
Any of various enzymatically catalyzed post-translational modifications of PEPTIDES or PROTEINS in the cell of origin. These modifications include carboxylation; HYDROXYLATION; ACETYLATION; PHOSPHORYLATION; METHYLATION; GLYCOSYLATION; ubiquitination; oxidation; proteolysis; and crosslinking and result in changes in molecular weight and electrophoretic motility.
Sequence Homology, Nucleic Acid
Protein Structure, Quaternary
Nucleic Acid Conformation
Enzymes that recombine DNA segments by a process which involves the formation of a synapse between two DNA helices, the cleavage of single strands from each DNA helix and the ligation of a DNA strand from one DNA helix to the other. The resulting DNA structure is called a Holliday junction which can be resolved by DNA REPLICATION or by HOLLIDAY JUNCTION RESOLVASES.
Saccharomyces cerevisiae Proteins
Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin.
A type of mutation in which a number of NUCLEOTIDES deleted from or inserted into a protein coding sequence is not divisible by three, thereby causing an alteration in the READING FRAMES of the entire coding sequence downstream of the mutation. These mutations may be induced by certain types of MUTAGENS or may occur spontaneously.
Highly reactive chemicals that introduce alkyl radicals into biologically active molecules and thereby prevent their proper functioning. Many are used as antineoplastic agents, but most are very toxic, with carcinogenic, mutagenic, teratogenic, and immunosuppressant actions. They have also been used as components in poison gases.
Open Reading Frames
A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).
An enzyme that catalyzes the conversion of 5-phosphoribosyl-1-pyrophosphate and hypoxanthine, guanine, or 6-mercaptopurine to the corresponding 5'-mononucleotides and pyrophosphate. The enzyme is important in purine biosynthesis as well as central nervous system functions. Complete lack of enzyme activity is associated with the LESCH-NYHAN SYNDROME, while partial deficiency results in overproduction of uric acid. EC 126.96.36.199.
Gene Expression Regulation
Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9)
Magnetic Resonance Spectroscopy
A rigorously mathematical analysis of energy relationships (heat, work, temperature, and equilibrium). It describes systems whose states are determined by thermal parameters, such as temperature, in addition to mechanical and electromagnetic parameters. (From Hawley's Condensed Chemical Dictionary, 12th ed)
Structural Homology, Protein
Membrane Transport Proteins
Mutation process that restores the wild-type PHENOTYPE in an organism possessing a mutationally altered GENOTYPE. The second "suppressor" mutation may be on a different gene, on the same gene but located at a distance from the site of the primary mutation, or in extrachromosomal genes (EXTRACHROMOSOMAL INHERITANCE).
Protein Interaction Domains and Motifs
Analysis of PEPTIDES that are generated from the digestion or fragmentation of a protein or mixture of PROTEINS, by ELECTROPHORESIS; CHROMATOGRAPHY; or MASS SPECTROMETRY. The resulting peptide fingerprints are analyzed for a variety of purposes including the identification of the proteins in a sample, GENETIC POLYMORPHISMS, patterns of gene expression, and patterns diagnostic for diseases.
Genes whose loss of function or gain of function MUTATION leads to the death of the carrier prior to maturity. They may be essential genes (GENES, ESSENTIAL) required for viability, or genes which cause a block of function of an essential gene at a time when the essential gene function is required for viability.
A set of genes descended by duplication and variation from some ancestral gene. Such genes may be clustered together on the same chromosome or dispersed on different chromosomes. Examples of multigene families include those that encode the hemoglobins, immunoglobulins, histocompatibility antigens, actins, tubulins, keratins, collagens, heat shock proteins, salivary glue proteins, chorion proteins, cuticle proteins, yolk proteins, and phaseolins, as well as histones, ribosomal RNA, and transfer RNA genes. The latter three are examples of reiterated genes, where hundreds of identical genes are present in a tandem array. (King & Stanfield, A Dictionary of Genetics, 4th ed)
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.
Hydrophobic and Hydrophilic Interactions
Bacterial Outer Membrane Proteins
Binding of the G domains of laminin alpha1 and alpha2 chains and perlecan to heparin, sulfatides, alpha-dystroglycan and several extracellular matrix proteins. (1/31010)The C-terminal G domain of the mouse laminin alpha2 chain consists of five lamin-type G domain (LG) modules (alpha2LG1 to alpha2LG5) and was obtained as several recombinant fragments, corresponding to either individual modules or the tandem arrays alpha2LG1-3 and alpha2LG4-5. These fragments were compared with similar modules from the laminin alpha1 chain and from the C-terminal region of perlecan (PGV) in several binding studies. Major heparin-binding sites were located on the two tandem fragments and the individual alpha2LG1, alpha2LG3 and alpha2LG5 modules. The binding epitope on alpha2LG5 could be localized to a cluster of lysines by site-directed mutagenesis. In the alpha1 chain, however, strong heparin binding was found on alpha1LG4 and not on alpha1LG5. Binding to sulfatides correlated to heparin binding in most but not all cases. Fragments alpha2LG1-3 and alpha2LG4-5 also bound to fibulin-1, fibulin-2 and nidogen-2 with Kd = 13-150 nM. Both tandem fragments, but not the individual modules, bound strongly to alpha-dystroglycan and this interaction was abolished by EDTA but not by high concentrations of heparin and NaCl. The binding of perlecan fragment PGV to alpha-dystroglycan was even stronger and was also not sensitive to heparin. This demonstrated similar binding repertoires for the LG modules of three basement membrane proteins involved in cell-matrix interactions and supramolecular assembly. (+info)
Arrestin function in G protein-coupled receptor endocytosis requires phosphoinositide binding. (2/31010)Internalization of agonist-activated G protein-coupled receptors is mediated by non-visual arrestins, which also bind to clathrin and are therefore thought to act as adaptors in the endocytosis process. Phosphoinositides have been implicated in the regulation of intracellular receptor trafficking, and are known to bind to other coat components including AP-2, AP180 and COPI coatomer. Given these observations, we explored the possibility that phosphoinositides play a role in arrestin's function as an adaptor. High-affinity binding sites for phosphoinositides in beta-arrestin (arrestin2) and arrestin3 (beta-arrestin2) were identified, and dissimilar effects of phosphoinositide and inositol phosphate on arrestin interactions with clathrin and receptor were characterized. Alteration of three basic residues in arrestin3 abolished phosphoinositide binding with complete retention of clathrin and receptor binding. Unlike native protein, upon agonist activation, this mutant arrestin3 expressed in COS1 cells neither supported beta2-adrenergic receptor internalization nor did it concentrate in coated pits, although it was recruited to the plasma membrane. These findings indicate that phosphoinositide binding plays a critical regulatory role in delivery of the receptor-arrestin complex to coated pits, perhaps by providing, with activated receptor, a multi-point attachment of arrestin to the plasma membrane. (+info)
Regulation of p53 function and stability by phosphorylation. (3/31010)The p53 tumor suppressor protein can be phosphorylated at several sites within the N- and C-terminal domains, and several protein kinases have been shown to phosphorylate p53 in vitro. In this study, we examined the activity of p53 proteins with combined mutations at all of the reported N-terminal phosphorylation sites (p53N-term), all of the C-terminal phosphorylation sites (p53C-term), or all of the phosphorylation sites together (p53N/C-term). Each of these mutant proteins retained transcriptional transactivation functions, indicating that phosphorylation is not essential for this activity of p53, although a subtle contribution of the C-terminal phosphorylation sites to the activation of expression of the endogenous p21(Waf1/Cip1)-encoding gene was detected. Mutation of the phosphorylation sites to alanine did not affect the sensitivity of p53 to binding to or degradation by Mdm2, although alteration of residues 15 and 37 to aspartic acid, which could mimic phosphorylation, resulted in a slight resistance to Mdm2-mediated degradation, consistent with recent reports that phosphorylation at these sites inhibits the p53-Mdm2 interaction. However, expression of the phosphorylation site mutant proteins in both wild-type p53-expressing and p53-null lines showed that all of the mutant proteins retained the ability to be stabilized following DNA damage. This indicates that phosphorylation is not essential for DNA damage-induced stabilization of p53, although phosphorylation could clearly contribute to p53 stabilization under some conditions. (+info)
Phosphorylation of the cap-binding protein eukaryotic translation initiation factor 4E by protein kinase Mnk1 in vivo. (4/31010)Eukaryotic translation initiation factor 4E (eIF4E) binds to the mRNA 5' cap and brings the mRNA into a complex with other protein synthesis initiation factors and ribosomes. The activity of mammalian eIF4E is important for the translation of capped mRNAs and is thought to be regulated by two mechanisms. First, eIF4E is sequestered by binding proteins, such as 4EBP1, in quiescent cells. Mitogens induce the release of eIF4E by stimulating the phosphorylation of 4EBP1. Second, mitogens and stresses induce the phosphorylation of eIF4E at Ser 209, increasing the affinity of eIF4E for capped mRNA and for an associated scaffolding protein, eIF4G. We previously showed that a mitogen- and stress-activated kinase, Mnk1, phosphorylates eIF4E in vitro at the physiological site. Here we show that Mnk1 regulates eIF4E phosphorylation in vivo. Mnk1 binds directly to eIF4G and copurifies with eIF4G and eIF4E. We identified activating phosphorylation sites in Mnk1 and developed dominant-negative and activated mutants. Expression of dominant-negative Mnk1 reduces mitogen-induced eIF4E phosphorylation, while expression of activated Mnk1 increases basal eIF4E phosphorylation. Activated mutant Mnk1 also induces extensive phosphorylation of eIF4E in cells overexpressing 4EBP1. This suggests that phosphorylation of eIF4E is catalyzed by Mnk1 or a very similar kinase in cells and is independent of other mitogenic signals that release eIF4E from 4EBP1. (+info)
Functional studies by site-directed mutagenesis on the role of Sp1 in the expression of the pyruvate kinase M and aldolase A genes. (5/31010)During the cell cycle of mitogen stimulated rat thymocytes, an 8-10-fold induction of glycolytic enzymes and a corresponding increase in the mRNA levels has been observed. This prompted us to study the transcriptional regulation of the rat aldolase A and pyruvate kinase M genes. cis-Regulatory elements of both promoters were evaluated by site-directed mutagenesis of promoter/luciferase constructs and transient transfections of rat hepatoma FTO2B cells. Furthermore, the binding proteins were identified by mobility shift assays in the presence of specific antibodies. In the aldolase AH1 promoter, five binding sites for Sp1 and Sp3 and a TPA responsive element were identified as essential for transcriptional regulation. Most of the promoter activity can be attributed to these regulatory elements. In the pyruvate kinase M promoter three out of five binding sites of Sp1 and Sp3 (B box and GC boxes 1 and 3) turned out to be functional in the transfection assays whereas the disruption of GC box 2 had no effect, and the disruption of the GC box 4 had only a minor effect on the promoter activity. Both promoters are stimulated by Sp1 as well as Sp3, as judged by cotransfection experiments of Drosophila SL2 cells. Therefore, the Sp1- and Sp3-directed transcription provides a means for common regulatory mechanism of the aldolase A and the pyruvate kinase M genes. (+info)
An antiviral mechanism of nitric oxide: inhibition of a viral protease. (6/31010)Although nitric oxide (NO) kills or inhibits the replication of a variety of intracellular pathogens, the antimicrobial mechanisms of NO are unknown. Here, we identify a viral protease as a target of NO. The life cycle of many viruses depends upon viral proteases that cleave viral polyproteins into individual polypeptides. NO inactivates the Coxsackievirus protease 3C, an enzyme necessary for the replication of Coxsackievirus. NO S-nitrosylates the cysteine residue in the active site of protease 3C, inhibiting protease activity and interrupting the viral life cycle. Substituting a serine residue for the active site cysteine renders protease 3C resistant to NO inhibition. Since cysteine proteases are critical for virulence or replication of many viruses, bacteria, and parasites, S-nitrosylation of pathogen cysteine proteases may be a general mechanism of antimicrobial host defenses. (+info)
Molecular dynamics of the sodium channel pore vary with gating: interactions between P-segment motions and inactivation. (7/31010)Disulfide trapping studies have revealed that the pore-lining (P) segments of voltage-dependent sodium channels undergo sizable motions on a subsecond time scale. Such motions of the pore may be necessary for selective ion translocation. Although traditionally viewed as separable properties, gating and permeation are now known to interact extensively in various classes of channels. We have investigated the interaction of pore motions and voltage-dependent gating in micro1 sodium channels engineered to contain two cysteines within the P segments. Rates of catalyzed internal disulfide formation (kSS) were measured in K1237C+W1531C mutant channels expressed in oocytes. During repetitive voltage-clamp depolarizations, increasing the pulse duration had biphasic effects on the kSS, which first increased to a maximum at 200 msec and then decreased with longer depolarizations. This result suggested that occupancy of an intermediate inactivation state (IM) facilitates pore motions. Consistent with the known antagonism between alkali metals and a component of slow inactivation, kSS varied inversely with external [Na+]o. We examined the converse relationship, namely the effect of pore flexibility on gating, by measuring recovery from inactivation in Y401C+E758C (YC/EC) channels. Under oxidative conditions, recovery from inactivation was slower than in a reduced environment in which the spontaneous YC/EC cross-link is disrupted. The most prominent effects were slowing of a component with intermediate recovery kinetics, with diminution of its relative amplitude. We conclude that occupancy of an intermediate inactivation state facilitates motions of the P segments; conversely, flexibility of the P segments alters an intermediate component of inactivation. (+info)
Functional consequences of mutations in the human alpha1A calcium channel subunit linked to familial hemiplegic migraine. (8/31010)Mutations in alpha1A, the pore-forming subunit of P/Q-type calcium channels, are linked to several human diseases, including familial hemiplegic migraine (FHM). We introduced the four missense mutations linked to FHM into human alpha1A-2 subunits and investigated their functional consequences after expression in human embryonic kidney 293 cells. By combining single-channel and whole-cell patch-clamp recordings, we show that all four mutations affect both the biophysical properties and the density of functional channels. Mutation R192Q in the S4 segment of domain I increased the density of functional P/Q-type channels and their open probability. Mutation T666M in the pore loop of domain II decreased both the density of functional channels and their unitary conductance (from 20 to 11 pS). Mutations V714A and I1815L in the S6 segments of domains II and IV shifted the voltage range of activation toward more negative voltages, increased both the open probability and the rate of recovery from inactivation, and decreased the density of functional channels. Mutation V714A decreased the single-channel conductance to 16 pS. Strikingly, the reduction in single-channel conductance induced by mutations T666M and V714A was not observed in some patches or periods of activity, suggesting that the abnormal channel may switch on and off, perhaps depending on some unknown factor. Our data show that the FHM mutations can lead to both gain- and loss-of-function of human P/Q-type calcium channels. (+info)
GENETAILOR SITE DIRECTED MUTAGENESIS PDF
GENETAILOR SITE DIRECTED MUTAGENESIS PDF - Site-specific mutagenesis techniques, also known as site-directed mutagenesis .. GeneTailor Site-Directed mutagenesis system (Invitrogen. I have designed
Analysis by Site-Directed Mutagenesis of Important Residues Invoved in the Assembly of Escherichia-Coli Heart-Labile NALYSIS BY...
Webb, Helen M. and Sixma, T.K. and Hol, W.G.J. and Hirst, Timothy R. (1994) Analysis by Site-Directed Mutagenesis of Important Residues Invoved in the Assembly of Escherichia-Coli Heart-Labile NALYSIS BY SITE-DIRECTED MUTAGENESIS OF IMPORTANT RESIDUES Enterotoxin. In: 6th European Workshop on Bacterial Protein Toxins, Stirling, Scotland. (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) ...
Mutagenesis Studies on the Membrane Anchoring Properties of Human CD2 - Enlighten: Theses
There is a lack of information about structural requirements for a functional transmembrane domain in eukaryotic membrane proteins. This led me to design a eukaryotic system to bring more information about this structure and the role of the positively charged residues situated at the cytoplasmic side of a transmembrane region. The CD2 molecule was chosen as a model for an integral membrane protein. I deleted the transmembrane domain (26 amino acids) by oligonucleotide site-directed mutagenesis and overlap extension using PCR mutagenesis. Truncated forms with transmembrane regions 14, 12, 10 and 8 amino acids long were created. The common positive cluster (Lys-Arg-Lys-Lys) at the cytoplasmic domain was disrupted by substituting it for polar residues (Gln-Gln-Gln-Gln). The effects of such mutagenesis was examined after expression of the mutant proteins in eukaryotic cells (COS-7 and CHO) . Cellular localization was determined by panning and immunostaining experiments. The functional state of the ...
Understanding the role of the essential Asp251 in cytochrome P450cam using site-directed mutagenesis, crystallography, and...
Fingerprint Dive into the research topics of Understanding the role of the essential Asp251 in cytochrome P450cam using site-directed mutagenesis, crystallography, and kinetic solvent isotope effect. Together they form a unique fingerprint. ...
Site-directed Mutagenesis Libraries - Creative Biostructure
Creative Biostructure develops comprehensive protein evolution and engineering platform to generate gene diversification by site-directed mutagenesis libraries.
Describe how you would use site-directed mutagenesis to, Ask an Expert
what are the criteria for good primers in a pcr reaction?describe how you would use site-directed mutagenesis to change, Hire Biology Expert, Ask Academics Expert, Assignment Help, Homework Help, Textbooks Solutions
Gene site saturated mutagenesis
Biomigas site-saturation mutagenesis technology systematically substitutes wildtype amino acids with partial or all 19 non-wildtype mutants.. Advantages. ...
The ion selectivity of the Na+ channel is determined by a ring of amino acids created by the P-loops of all four domains (D1-D4), and the major determinants are the residues referred to as `DEKA (Asp372, Glu898, Lys1419, and Ala1711; following residue numbers are the equivalents of human Nav1.5) (Heinemann et al., 1992; Sun et al., 1997) (Fig. 6A). However, the contribution of each P-loop to the ion selectivity is not equal (Schlief et al., 1996). Site-directed mutagenesis studies have shown that the positive Lys residue in D3 is the most critical residue for ion selectivity (Chiamvimonvat et al., 1996). In contrast, the physiological role of the D4 selectivity filter region is less clear. Cysteine mutagenesis experiments have shown that the five residues between Ser1710 and Asp1714 around the selectivity filter residue Ala1711 are all accessible from outside (Chiamvimonvat et al., 1996), suggesting that these five residues face the hydrophilic ion-conducting pore. In fact, mutations at the ...
Amino acids within TMs have been shown to play important roles in substrate binding, maintenance of protein stability, and correct folding of proteins (Hong et al., 2004, 2010; Gui and Hagenbuch, 2009; Miyagawa et al., 2009; Li et al., 2012). Studies of single nucleotide polymorphisms have also pointed out that mutants located within TMs often result in functional changes (Kalliokoski and Niemi, 2009). In the present study, we used site-directed mutagenesis to study the involvement of amino acid residues within the putative TM6 of OATP1B1 in substrate transport. Of the 24 amino acids analyzed, F262A, V264A, L267A, S269A, F276A, and F278A showed a more than 60% reduction of transport activity. Further studies revealed that these mutants had much reduced transporter protein expression compared with the wild-type. If protein expression is considered in the comparison of the transport activity, E-3-S uptake by these mutants was more than 60% of that by wild-type OATP1B1 (data not shown). Therefore, ...
Mutation of the ATP-Binding Pocket of SSA1 Indicates That a Functional Interaction Between Ssa1p and Ydj1p Is Required for Post...
Site-directed mutagenesis, cloning, strains, and media: Four mutant alleles of SSA1, ssa1-101 (K69Q), ssa1-102 (G198D), ssa1-103 (G199D), and ssa1-104 (G226D), were created using the Quick-Change site-directed mutagenesis kit (Stratagene, La Jolla, CA). SSA1 inserted between the HindIII and BamHI sites of YEp351 (Hillet al. 1986; YEp351-SSA1 plasmid provided by E. Craig, University of Wisconsin, Madison) served as the template. The primer pairs used were as follows: for ssa1-101, 5′-CCGTTTTCGACGCTCAGCGTTTGATCGG-3′ and 5′-CCGATCAAACGCTGAGCGTCGAAAACGG-3′; for ssa1-102, 5′-GTCTTGATTTTCGACTTGGATGGTGGTACTTTCG-3′ and 5′-CGAAAGTACCACCATCCAAGTCGAAAATCAA GAC-3′; for ssa1-103,5′-GATTTTCGACTTGGGTGATGG TACTTTC GATGTC-3′ and 5′-GACATCGAAAGTACCATCACCCAAGTC GAAAATC-3′; and for ssa1-104, 5′-GGTGACACCCATTTGGAT GGTGAAGATTTTGAC-3′ and 5′-GTCAAAATCTTCACCATC CAAATGGGTGTCACC-3′. In each primer the altered nucleotide is boldfaced and underlined. Mutagenesis resulted in plasmids ...
Site-directed mutagenesis - experimental considerations | NEB
Site-drected mutagenesis considerations include: primer design, modification and purity, template removal, ligation, transformation, and product evaluation.
Lidstrom:Site-Directed Mutagenesis - OpenWetWare
Our lab (at least Amanda and Janet) have not tried it yet. Our understanding is that it is for more substantial changes than just single amino acid changes ...
Wikiomics:Site Directed Mutagenesis - OpenWetWare
This page is beta. Please feel free to update it. The idea here is to create a flow of work to simulate the point mutation of a single amino acid in a protein. ...
SDM-Assist software to design site-directed mutagenesis primers introducing silent restriction sites
The direct identification of SDM clones will save time and money for researchers. SDM-Assist also scores the primers based on factors such as Tm, GC content and secondary structure allowing for simplified selection of optimal primer pairs.
Site directed mutagenesis
you will bite yourself in the a** when you later discover that there was a mutation somewhere else in your sequence. Considering the cost of commercial sequencing (the cheapest way are those packages where they just run the reaction and you get the raw data and do the evaluation by yourself, there is free software available like finch tv), sequencing your constructs is a must before you derive any data from them that you want to publish/patent or which are important in any other context. I had learned my lesson once by finding out almost too late that one of the primers I had used was missing a base (still in the time of running your own DNA synthesizer and sequencing with gels and 35S) my 2c Wo On 15 Aug., 04:05, mnr mnr ,mnr... from gmail.com, wrote: , If I were to go down the Phusion road, will I need to sequence the entire of , my gene (4.5 kbp) or just the mutated region to make sure the protein , finally has mutated in the right places? ...
Site Directed Mutagenesis - 100% Präzision
Ortsgerichtete Mutagenese: Punktmutationen, Aminosäure-Austausche und Insertionen einführen. Bis zu 150 bp. Kurze Lieferzeit. Absolute Vertraulichkeit.
Mutagenesis: Site-specific - eLS - El-Gewely - Wiley Online Library
Site-specific mutagenesis techniques are aimed at the precise substitution, insertion or deletion of any coding sequence in vitro. More recently, however, such precise alterations are also being developed for in vivo gene/genome modifications. These techniques are revolutionizing our understanding of the genetic and molecular mechanisms in several biological systems, which could lead to the development of new enzymes, therapeutics as well as improved agricultural applications. ...
Ergebnisse durchsuchen | Europäische Behörde für
Deadline: 4 June 2020 EFSAs GMO Panel has launched an open consultation on its draft scientific opinion on the applicability of the EFSA opinion on site-directed nucleases type 3 (SDN-3) for the safety assessment of plants developed using site-dire .... ...
Altmetric - Site-Directed Mutagenesis Study on the Thermal Stability of a Chimeric PQQ Glucose Dehydrogenase and Its Structural...
Older research outputs will score higher simply because theyve had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 271,050 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 61% of its contemporaries ...
Structural and functional analysis of the canine histamine H2 receptor by site-directed mutagenesis: N-glycosylation is not...
G-protein-coupled receptors generally share a similar structure containing seven membrane-spanning domains and extracellular site(s) for N-glycosylation. The histamine H2 receptor is a member of the family of G-protein-coupled receptors, and has three extracellular potential sites for N-glycosylation (Asn-4, Asn-162 and Asn-168). To date, however, no information has been presented regarding N-glycosylation of the H2 receptor. To investigate the presence, location and functional roles of N-glycosylation of the H2 receptor, site-directed mutagenesis was performed to eliminate the potential site(s) for N-glycosylation singly and collectively. The wild-type and mutated H2 receptors were expressed stably in Chinese hamster ovary (CHO) cells or transiently in COS7 cells. Immunoblotting of the wild-type and mutated H2 receptors with an antiserum directed against the C-terminus of the H2 receptor showed that mutation at Asn-162, but not at Asn-168, resulted in a substantial decrease in the molecular ...
Site-directed mutagenesis of the active site of diacylglycerol kinase α: calcium and phosphatidylserine stimulate enzyme...
Diacylglycerol kinases (DAGKs) catalyse ATP-dependent phosphorylation of sn-1,2-diacylglycerol that arises during stimulated phosphatidylinositol turnover. DAGKα is activated in vitro by Ca2+ and by acidic phospholipids. The regulatory region of DAGKα includes an N-terminal RVH motif and EF hands that mediate Ca2+-dependent activation. DAGKα also contains tandem C1 protein kinase C homology domains. We utilized yeast, Saccharomyces cerevisiae, which lacks an endogenous DAGK, to express DAGKα and to determine the enzymic activities of different mutant forms of pig DAGKα in vitro. Six aspartate residues conserved in all DAGKs were individually examined by site-directed mutagenesis. Five of these aspartate residues reside in conserved blocks that correspond to sequences in the catalytic site of phosphofructokinases. Mutation of D434 (Asp434) or D650 abolished all DAGKα activity, whereas substitution of one among D465, D497, D529 and D697 decreased the activity to 6% or less of that for ...
Site-Directed Mutagenesis - Cloning and Mutagenesis - GENEWIZ
GENEWIZ can increase your research productivity by performing your time-consuming site-directed mutagenesis projects efficiently and cost-effectively. Our customized mutagenesis services provide a fail-safe approach to obtain mutant constructs quickly, with 100% accuracy, thus eliminating the possibility of undesired mutations in your gene.. ...
Plasmid-based one-pot saturation mutagenesis<...
TY - JOUR. T1 - Plasmid-based one-pot saturation mutagenesis. AU - Wrenbeck,Emily E.. AU - Klesmith,Justin R.. AU - Stapleton,James A.. AU - Adeniran,Adebola. AU - Tyo,Keith E.J.. AU - Whitehead,Timothy A.. PY - 2016/11/1. Y1 - 2016/11/1. N2 - Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped plasmid dsDNA. The method can be used to produce comprehensive or single- or multi-site saturation mutagenesis libraries.. AB - Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Here we present nicking mutagenesis, a robust, ...
The limitations of site-directed mutagenesis in the localization of Rh D epitopes | Chang, Tylis Y.; Siegel, Don L. | download
The limitations of site-directed mutagenesis in the localization of Rh D epitopes | Chang, Tylis Y.; Siegel, Don L. | download | BookSC. Download books for free. Find books
Defining fibronectins cell adhesion synergy site by site-directed mutagenesis. | [email protected]
Defining fibronectins cell adhesion synergy site by site-directed mutagenesis.s profile, publications, research topics, and co-authors
OneClick: A Program for Designing Focused Mutagenesis Experiments
OneClick is a user-friendly web-based program, developed specifically for quick-and-easy design of focused mutagenesis experiments (|em|e.g|/em|., site-directed mutagenesis and saturation mutagenesis). Written in Perl and developed into a web application using CGI programming, OneClick offers a step-by-step experimental design, from mutagenic primer design to analysis of a mutant library. Upon input of a DNA sequence encoding the protein of interest, OneClick designs the mutagenic primers according to user input, |em|e.g|/em|., amino acid position to mutate, type of amino acid substitutions (|em|e.g|/em|., substitution to a group of amino acids with similar chemical property) and type of mutagenic primers. OneClick has incorporated an extensive range of commercially available plasmids and DNA polymerases suitable for focused mutagenesis. Therefore, OneClick also provides information on PCR mixture preparation, thermal cycling condition, expected size of PCR product and agar plate to use during bacterial
A Periplasmic Drug-Binding Site of the AcrB Multidrug Efflux Pump: a Crystallographic and Site-Directed Mutagenesis Study |...
The drug binding also produces significant displacement of several segments of the backbone. There is a large movement of the segment between Val107 and Leu113 (including the mutated residue Ala109), apparently because the segment between Gln104 and Gln108 becomes helical as a result of drug binding. Some of the side chain atoms appear to become displaced by nearly 3 Å upon drug binding. In addition, the segment between Gly861 and Gly870, just preceding TM helix 8, shows a slight backbone displacement and side chain movement up to 3 Å.. Similar interactions often involving the same set of residues appear to occur with the binding of R6G and Et in our model (not shown). Surprisingly, no acidic residue that would neutralize the positive charges of these dyes was found within 6 Å of the ligand (except Asp566, which is close to the benzoic acid moiety of R6G but more than 11 Å away from its amine nitrogens), although the partially negative π-electron cloud of the phenyl ring of Phe664 is about ...
Homemade Site Directed Mutagenesis of Whole Plasmids | Protocol
Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. Here we...
Structure Cluster - 1RQL: Crystal Structure of Phosponoacetaldehyde Hydrolase Complexed with Magnesium and the...
1RQL: X-ray crystallographic and site-directed mutagenesis analysis of the mechanism of Schiff-base formation in phosphonoacetaldehyde hydrolase catalysis
Mutagenesis | BaseClear B.V.
Site-directed mutagenesis of a sequence to make specific, targeted changes to double stranded DNA. Based on optimised protocols, BaseGene is able to create mutations in virtually every DNA fragment. This is a valuable tool for studying DNA or protein structure and function. Site-directed or random mutagenesis can be applied to any DNA-fragment cloned into a plasmid vector. We can help you to introduce a range of mutations, varying from point mutations to deletions or insertions.. ...
Indian Patents. 247720:PROCESS FOR GENERATING AND DETECTING RECOMBINANT DNA SEQUENCES IN PROKARYOTES
Directed evolution has so far been used almost exclusively as a tool for engineering proteins. By mutation techniques such as site-directed mutagenesis, cassette-mutagenesis, random mutagenesis, and error prone PCR variants of protein functions have been generated and the libraries thus produced have been screened for their ability to perform a specific function. Recursive application of this procedure has been used successfully for the modification of physical and catalytic properties of enzymes such as pH optima, thermo-tolerance, solvent stability, stereoselectivity, catalytic activity and substrate specificity as well as toxicity resistance mechanisms in bacteria and the host range and stability of viruses.Traditional mutagenesis approaches for evolving new properties in enzymes have a number of limitations. These approaches are only applicable to genes or sequences that have been cloned and functionally characterized and that have a discrete function. Also, the traditional mutagenesis ...
Q5 Site-Directed Mutagenesis Kit - Lab-JOT® Ltd. Sp.z o.o. Sp.k.
Site Directed Mutagenesis with Pfu Turbo
Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol ...
mutagenesis expert come in~ - Molecular Cloning - BioForum
mutagenesis expert come in~ - posted in Molecular Cloning: I want to do a site directed mutagenesis in a 6kb plasmid, the primer set is primerF: GCCTCTGCTTAGTGGaaTTcCCACCCCCACAACCCGCAGG primerR: CCTGCGGGTTGTGGGGGTGGgAAttCCACTAAGCAGAGGC, The reaction system was designed as following: template 1ul (100ng) primer 1+1 ul (10mM each) dNTPs 1 ul (25mM each) 10Xpfu buffer 5 ul pfu 1 ul H2O...
Faculty Profile of J. Bryan Jones
G. DeSantis and J.B. Jones, Combining Site-Specific Chemical Modification with Site-Directed Mutagenesis: A Versatile Strategy To Move Beyond the Structural Limitations of the 20 Natural Amino Acids Side Chains in Protein Engineering , in In Vitro Mutagenesis Protocols , Methods in Molecular Biology, Humana Press, 2002, vol 182, pp55-65 ...
Weird problem in in vitro mutagenesis: post #1
Weird problem in in vitro mutagenesis - posted in Molecular Biology: Hi, Iam doing invitro mutagenesis for a 5.2Kb plasmid using stratagene kit. I did not get any results with my plasmid, so checked the control in the kit. Unfortunately the control is not working. I have tried many times, but got no colonies. The conditions used were according to the instructions given by the manufacturer. 95 C 50 sec 95 C 50 seconds 55 C 1 min 68 C 5 minute x 18 times 68 C 5 minutes. I even tried addin...
The National Institute of General Medical Sciences (NIGMS) has a strong track record of funding scientists who receive a Nobel Prize. The Nobel Prize was created by Swedish inventor Alfred Nobel. The international award has been given yearly since 1901 for achievements in physics, chemistry, physiology or medicine, literature, and peace. Another category, economics, was added by the Nobel Foundation in 1968. Winners receive their awards on December 10, the anniversary of Nobels death. For more facts about the Nobel Prize, visit https://www.nobelprize.org/.. Since its creation in 1962, NIGMS has supported the work of 90 Nobel laureates-43 in physiology or medicine and 47 in chemistry. These investigators perform cutting-edge basic research that is the foundation for understanding normal life processes and disease. Such important breakthroughs in chemistry and biology often lead to more focused research that, years later, leads to important medical advances or products such as medicines or ...
Protocols and Video Articles Authored by Kajohn Boonrod
Kajohn Boonrod is the author of this article in the Journal of Visualized Experiments: Homemade Site Directed Mutagenesis of Whole Plasmids
Team:Warsaw/Calendar-Stage1/14 September 2010 - 2010.igem.org
Directed mutagenesis was preformed on pSB-MinC in order to remove an additional PstI site. Two primers were used: MinCmutF (TAATTGCAGTCGCGCCGC) and MinCmutR (GTCGAAAACGCTTTGACCG). PCR was run with Yellow Pfu Polimerase (Eurex) in a standard reaction mix ...
New insights into the structural features and functional relevance of human cytochrome P450 2C9. Part II<...
TY - JOUR. T1 - New insights into the structural features and functional relevance of human cytochrome P450 2C9. Part II. AU - Mo, Sui Lin. AU - Zhou, Zhi Wei. AU - Yang, Li Ping. AU - Wei, Ming Qian. AU - Zhou, Shu Feng. PY - 2009/12. Y1 - 2009/12. N2 - Part I of this article published in the same issue of Current Drug Metabolism discussed the substrate specificity, inhibitor selectivity and structure-activity relationship (SAR) of human CYP2C9. The features of CYP2C9 pharmacophore and SAR models have been elaborated. Part II of this article will address the homology models of CYP2C9, data from site-directed mutagenesis studies, and crystal structural features of CYP2C9. The heteroactivation of CYP2C9 and its interactions with other CYPs will also be discussed. A number of ligand-based and homology models of CYP2C9 have been reported and this has provided insights into the binding of ligands to the active site of CYP2C9. Site-directed mutagenesis studies have revealed that a number of residues ...
The coordination sphere of iron-sulfur clusters: lessons from site-directed mutagenesis experiments | SpringerLink
Cysteine is the ubiquitous ligand of iron-sulfur clusters in proteins, although chemical models have indicated that functional groups other than thiolates can coordinate iron in iron-sulfur...
Identification of Critical Amino Acid Residues in Human Immuno...
Identification of Critical Amino Acid Residues in Human Immunodeficiency Virus Type 1 IN Required for Efficient Proviral DNA Formation at Steps prior to Integra
The structure of tumour necrosis factor--implications for biological function. - CAMS Oxford Institute
The three-dimensional structure of TNF has been determined at 0.29 nm using the technique of X-ray crystallography. Published data on site-directed mutagenesis and antibody binding may now be assessed in the light of the structure, thus the links between structure and function for TNF may be addressed. TNF is a compact trimer composed of three identical subunits of 157 amino acids. The main-chain topology for a single subunit is essentially a beta-sandwich structure formed by two anti-parallel beta-pleated sheets. This mainchain fold corresponds to the jelly roll motif observed in viral coat proteins such as VP1, VP2 and VP3 of rhinovirus, or the hemagglutinin molecule of influenza. TNF is the first non-viral protein to contain this motif. The subunits associate tightly about a threefold axis interacting through a simple edge-to-face packing of the beta-sandwich to form the solid, conical shaped trimer. A large number of the residues conserved between the amino acid sequences of TNF and lymphotoxin
SPO14 Separation-of-Function Mutations Define Unique Roles for Phospholipase D in Secretion and Cellular Differentiation in...
Plasmids ME1646 (S29, nucleotide 2555 A → G; amino acid 853 H → R), ME1647 (S41, nucleotide 2555 A → G; amino acid 853 H → R), and ME1648 (S55, nucleotides 2260 A → G, 2545 G → A; amino acids 754 N → D, 849 A → T) were sequenced between the DraI and HpaI sites to determine the mutations responsible for the mutant phenotypes. Site-directed mutagenesis was performed to generate the single (S29/S41) and double mutations (S55) using the QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA) as recommended. The primers used were: S29/S41 P126, 5′-CAG AAT TGC TGA TTT TCG TTT AGA TAA ACC ATT C, and P127, 5′-G AAT GGT TTA TCT AAA TCA CGA AAA TCA GCA ATT CTG; S55 (nucleotide 2260 A → G) P135, 5′-TTG TGA TTT ATA GGG ATG TGG GGA ATA TTG, and P136, 5′-CAA TAT TCC CCA CAT CCC TAT AAA TCA CAA; and S55 (nucleotide 2545 G → A) P132, 5′-TAT TCA AAT GCC AGA ATT ACT GAT TTT CAT GAT TTA G, and P133, 5′-C TAA ATC ATG AAA ATC AGT AAT TCT GGC ATT TGA ATA (the boldface ...
Examination of alternate codon bias solutions for expression and purification of recombinant mechano-growth factor in...
Mechano-growth factor (MGF), an alternative splicing variant of insulin-like growth factor-1 (IGF-1) gene, promotes cell proliferation and inhibits cell differentiation. It also plays an important role in tumor development. It is important to optimize the production process and achieve MGF protein because there is no commercial MGF protein available. In this study, the human MGF gene is cloned into pGEX-4T-1 and the recombinant human MGF (rhMGF) protein could be expressed in Rosetta (DE3) by isopropyl β-D-1-thiogalactopyranoside induction but not in BL21 (DE3). Mutation from rare codons to Escherichia coli preferred ones is performed. We obtain MGF(Mut54-56) and MGF(Mut-total) fragments through site-directed mutagenesis and overlapping PCR. Both pGEX-4T-1/MGF(Mut54-56)- and pGEX-4T-1/MGF(Mut-total)-transformed BL21 (DE3) can be induced to express rhMGF protein. To optimize the production technology, expression and purification of rhMGF are analyzed and compared in Rosetta (DE3) and BL21 (DE3). Results
UniProtKB/SwissProt variant VAR 026378
Cited for: VARIANTS TRP-18; CYS-34; HIS-34; LEU-216; SER-286; SER-291; MET-299; CYS-376; GLY-447; ALA-449; VAL-461; SER-475; TRP-481; TYR-524; ARG-558; HIS-568; ARG-579; LYS-592; GLY-596; ALA-601; PHE-618; ASP-638; LEU-656; THR-672; HIS-689; LYS-692; PHE-705; ILE-924; GLN-986; MET-1016; ARG-1040; ALA-1082; LEU-1090; LEU-1098; TYR-1103; LYS-1107; TRP-1116; GLN-1193; MET-1251; SER-1293; PHE-1308; TRP-1512; ASN-1787; THR-1836; LYS-1901; CYS-1919; LEU-1951; GLN-1958; LEU-1962; MET-1968; GLN-1991; LEU-2004 AND ALA-2006; VARIANTS BRGDA1 GLN-18; LYS-70; ASN-84; SER-93; SER-94; GLN-104; TRP-104; LYS-109; GLN-121; TRP-121; GLU-126; PRO-136; MET-146; GLN-161; LYS-161; ASN-175; GLY-178; ARG-182; VAL-185; VAL-204; GLN-212; ILE-220; GLN-222; LEU-223; TRP-225; VAL-226; ILE-232; MET-240; LYS-270; GLN-276; ASP-278; CYS-282; ILE-300; PRO-315; ASN-320; ARG-325; LEU-336; ASP-351; VAL-351; ASN-356; CYS-367; HIS-367; LEU-367; LYS-369; GLY-374; HIS-376; ARG-386; GLU-386; ALA-396; LEU-396; LYS-439; GLY-501; HIS-526; ...
TRANSIENT IMMORTALIZATION - Patent application
31150DNAArtificial Sequencesynthetic primer for the site-directed mutagenesis 1cctccccctg aacctgaaac aagatctgaa tgcaattgtt gttgttaacg 50251DNAArtificial Sequencesynthetic primer for the site-directed mutagenesis 2cgttaacaac aacaattgca ttcaggatct tgtttcaggt tcagggggag g 51316DNAArtificial Sequencesynthetic primer, RT-telo-4 3ctcatatatt cagtat 16415DNAArtificial Sequencesynthetic primer, RT-telo-3 4ctggacactc gctca 15515DNAArtificial Sequencesynthetic primer, RT-telo-2 5tcagccggac atgca 15615DNAArtificial Sequencesynthetic primer, RT-telo-1 6tcactcaggc ctcag 15717DNAArtificial Sequencesynthetic primer, RCR-telo-10 7gctggtgtct gctctcg 17825DNAArtificial Sequencesynthetic primer, RCR-telo-11 8ctgcagcagg aggatcttgt agatg 25921DNAArtificial Sequencesynthetic primer, RCR-telo-6 9gcaggtgaac agcctccaga c 211024DNAArtificial Sequencesynthetic primer, RCR-telo-R13 10cacaggctgc agagcagcgt ggag 241125DNAArtificial Sequencesynthetic primer, RCR-telo-F12 11gtcctacgtc cagtgccagg ggatc 251225DNAArtificial ...
Updated mutation overview - VU Medicinal Chemistry - Tools & Downloads
For our British Journal of Pharmacology publication regarding the structural chemogenomics analysis of aminergic GPCRs (publication) we created a comprehensive overview of all site-directed mutagenesis studies performed against HRH1, HRH2, HRH3, HRH4, DRD3, ACM2, ACM3, ADRB1 and ADRB2 (aminergic GPCRs). In total 1420 reported single-point mutations were registered (including their effect on the binding affinity of reported ligands) at 128 individual B&W positions. ...
UniProtKB/SwissProt variant VAR 073451
Cited for: VARIANTS EIEE6 PHE-17 DEL; THR-68; ASN-79; CYS-84; PRO-98; GLN-101; TRP-101; ARG-108; ASP-127; ARG-199; SER-227; THR-227; SER-232; ARG-233; VAL-342; ASP-343; TRP-351; SER-359; ARG-363; ARG-384; CYS-393; HIS-393; VAL-400; VAL-403; PHE-406; GLY-626; ASP-762; THR-785; ILE-812; ARG-842; 854-GLY-LEU-855 DEL; CYS-859; GLN-862; PRO-890; CYS-932; PRO-933; CYS-946; HIS-946; ARG-950; LYS-954; LYS-956; LEU-957; ILE-976; VAL-979; ARG-993; 999-ASN-LEU-1000 DELINS LEU-ILE-SER; LYS-1208; LYS-1221; PHE-1230; ASP-1238; ALA-1266; ASN-1288; VAL-1320; PRO-1326; GLY-1350; ARG-1358; PRO-1370; HIS-1378; THR-1378; ILE-1394; TYR-1396; SER-1417; PHE-1423; ALA-1429 DEL; VAL-1433; LYS-1450; SER-1451; LYS-1454; HIS-1462; LYS-1476; LYS-1503; GLY-1544; GLU-1586; ARG-1588; HIS-1592; PRO-1592; SER-1605; GLU-1637; THR-1638; CYS-1648; GLU-1653; PRO-1660; PRO-1667; LEU-1668; ILE-1672; THR-1673; THR-1683; ASP-1684; TRP-1688; ARG-1714; ASN-1763; ASN-1770; PHE-1770; THR-1770; THR-1780; VAL-1783; LYS-1787; PRO-1832; ...
JoVE Author Search: Br%C3%A9maud L
Sodium, Electrophoresis, Escherichia, Escherichia Coli, Family, Flexibility, Inhibition, Muscle, Mutagenesis, PH, Polyacrylamide Gel Electrophoresis, Polymerization, Polymers, Report, Serpins, Site-directed Mutagenesis, Skeletal Muscle, Sodium Dodecyl Sulfate, Temperature
We have constructed highly constitutively active H3 receptors (MT6; A357K) by induced point mutagenesis in the C-terminal portion of the third intracellular loop of mouse H3 receptors. This substitution resulted in three major biochemical modifications of the H3 receptors: 1) increased constitutive activity leading to agonist-independent inhibition of adenylyl cyclase in cells; 2) moderately increased affinity for some agonists; and 3) reduced affinities for antagonist acting as inverse agonists.. Several mutagenesis studies indicated that the C-terminal portion of the i3 might act in concert with other structural domains to determine the selectivity and efficiency of G-protein-coupling receptor. For the adrenergic receptors, it has been shown that the C-terminal portion of the i3 plays a crucial role in G-protein-coupling receptor (ODowd et al., 1988; Cotecchia et al., 1990; Liggett et al., 1991). Ren et al. (1993) previously showed that the T373K mutation adjacent to TM6 causes dramatically ...
Tryptophan scanning mutagenesis reveals distortions in the helical structure of the δM4 transmembrane domain of the Torpedo...
Caballero-Rivera, D., Cruz-Nieves, O.A., Oyola-Cintrón, J., Torres-Nunez, D.A., Otero-Cruz, J.D., Lasalde-Dominicci, J.A. (2012) Channels (Austin) 2012 Mar-Apr 6(2):111-23 ...
Mixed bases are used in primers to bind to templates that contain variability or a mixture of sequences at the primer binding sites. Mixed bases can also be used to create diversity in clone libraries and in site directed mutagenesis.. IDT offers random base oligos. Order them by using an upper case letter N or other IUPAC-IUB symbol (see table below). IDT offers two types of randomization, machine mix and hand mix. Machine mix bases are charged at the standard base price for the scale ordered. Hand mixing is done to provide custom ratios of the bases, and incurs an additional charge. When entering your sequence, the Mixed Base tab at the bottom of the page lists the IUB symbols and is where custom mix ratios need to be entered ...
In Vitro Mutagenesis Protocols (Methods... | WHSmith Books
Buy In Vitro Mutagenesis Protocols (Methods in Molecular Biology v. 57) by TROWER From WHSmith today! FREE delivery to store or FREE UK delivery on all ...
RCSB PDB - 1ENA: CRYSTAL STRUCTURES OF THE BINARY CA2+ AND PDTP COMPLEXES AND THE TERNARY COMPLEX OF THE ASP 21->GLU...
1ENA: Crystal structures of the binary Ca2+ and pdTp complexes and the ternary complex of the Asp21-->Glu mutant of staphylococcal nuclease. Implications for catalysis and ligand binding.
Noncysteinyl coordination to the [4Fe-4S]2+ cluster of the DNA repair adenine glycosylase MutY introduced via site-directed mutagenesis. Structural characterization of an unusual histidinyl-coordinated cluster ...
Dear Netters I am looking for the GUS gene from which the glycosylation sites have been removed by site directed mutagenesis. I know it is around but I dont have the reference. Sjef Smeekens Utrecht University email: wbtmcbw at cc.ruu.nl ...
pCDNA3.1-human-ub-wt | mBIOnet
Virtual plasmid consisting of wild-type human ubiquitin cloned between KpnI and NotI sites of pCDNA3.1(+)-Zeo. Used for site-directed mutagenesis tutorial.. 1 GACGGATCGG GAGATCTCCC GATCCCCTAT GGTCGACTCT ...
KRIBB: Search Results
Production of ser125-interleukin-2 by site directed mutagenesis / Seongman Kang; Ju Won Kwak; Jong Bum Kwon; Sung Wan Kim; In Young Lee; Sun Bok Lee; Hye-Young Yun; Kyung Soo Hahm; Moon Hi Han; Doe Sun Na , 1989 ...
An efficient one-step site-directed deletion, insertion, single and multiple-site plasmid mutagenesis protocol - Nuffield...
Mutagenesis of the mouse Bhlhb4 gene. A: Gene targeting | Open-i
Mutagenesis of the mouse Bhlhb4 gene. A: Gene targeting strategy showing partial restriction map of WT Bhlhb4 allele, the targeting vector, the targeted ES cell
Melet, A., Marques-Soares, C., Schoch, G. A., Macherey, A. C., Jaouen, M., Dansette, P. M., Sari, M. A., Johnson, E. F., Mansuy, D. Analysis of human cytochrome P4502C8 substrate specificity using a substrate pharmacophore and site-directed mutants Biochemistry 2004 43:15379-15392 DOI:10.1021/bi0489309 PMID:15581350 ...
Tools to Study Mammalian Mutagenesis - Stephen Dertinger
Despite the extensive work being performed to understand cancer and carcinogenic properties of chemicals and other agents, there are still gaps in our ability t...
Acid Resource | Learn About, Share and Discuss Acid At Popflock.com
Get Acid essential facts. View Videos or join the Acid discussion. Add Acid to your PopFlock.com topic list or share. Acid at popflock.com
Nobel Lecture on Invention of Site-Directed Mutagenesis OpenWetWare Diagram summarizing site-directed mutagenesis (CS1 maint: ... Direct gene deletion and site-specific mutagenesis with PCR and one recyclable marker Direct gene deletion and site-specific ... Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis, it is used for investigating the structure and ... "SDM-Assist software to design site-directed mutagenesis primers introducing "silent" restriction sites". BMC Bioinformatics. 14 ...
... is a type of site-directed mutagenesis that uses a short, double-stranded oligonucleotide sequence (gene ... Unlike many site directed mutagenesis methods, cassette mutagenesis also does not involve primer extension by DNA polymerase. ... mutagenesis, sequencing, expression) can be performed in the same plasmid. Worrall, Andrew (1994). "Site-Directed Mutagenesis ... Compared to mutagenesis methods that requires the synthesis of double stranded DNA using a single stranded template (1-30% in ...
... is commonly achieved by site-directed mutagenesis PCR with a randomised codon in the primers (e.g. SeSaM ... Site saturation mutagenesis (SSM), or simply site saturation, is a random mutagenesis technique used in protein engineering, in ... "An efficient one-step site-directed and site-saturation mutagenesis protocol". Nucleic Acids Research. 32 (14): e115. doi: ... Saturation mutagenesis is commonly used to generate variants for directed evolution. Sequence Saturation Mutagenesis Reetz, M. ...
Mutagenesis (molecular biology technique)
Site-directed mutagenesis has proved useful in situations that random mutagenesis is not. Other techniques of mutagenesis ... Site directed mutagenesis allows the effect of specific mutation to be investigated. There are numerous uses; for example, it ... Random mutagenesis cannot target specific regions or sequences of the genome; however, with the development of site-directed ... The experiment used site directed mutagenesis to mimic the expected mutations of the specific chemical. The mutation resulted ...
Directed mutagenesis Insertion (genetics) PCR mutagenesis Site-directed mutagenesis Transposon mutagenesis Biffi A, et al., ... Wikimedia Commons has media related to Insertional mutagenesis. Insertional+mutagenesis at the US National Library of Medicine ... Insertional mutagenesis is possible whether the virus is of the self-inactivating types commonly used in gene therapy or ... In molecular biology, insertional mutagenesis is the creation of mutations of DNA by addition of one or more base pairs. Such ...
Site-directed mutagenesis mapping. The molecular biological technique of site-directed mutagenesis (SDM) can be used to enable ... Shotgun mutagenesis is a high-throughput approach for mapping the epitopes of mAbs. The shotgun mutagenesis technique begins ... One of its advantages is that it determines the interaction site of the antigen-antibody complex in its native solution, and ... In immunology, epitope mapping is the process of experimentally identifying the binding site, or epitope, of an antibody on its ...
Biotin attachment domain
Analysis by site-directed mutagenesis". J. Biol. Chem. 267 (26): 18407-18412. doi:10.1016/S0021-9258(19)36977-7. PMID 1526981. ...
Prostaglandin DP1 receptor
Identification by site-directed mutagenesis". J. Biol. Chem. 275 (32): 24294-303. doi:10.1074/jbc.M002437200. PMID 10827082. ...
Identification by site-directed mutagenesis". The Journal of Biological Chemistry. 268 (6): 3980-5. doi:10.1016/S0021-9258(18) ... Catalytic properties, subunit structure, and localization of the gamma-glutamyl binding site on the light subunit". The Journal ...
Analysis using site-directed mutagenesis". The Journal of Biological Chemistry. 265 (34): 21027-31. PMID 2250008. Kim WS, ...
MacDonald NJ, Freije JM, Stracke ML, Manrow RE, Steeg PS (1996). "Site-directed mutagenesis of nm23-H1. Mutation of proline 96 ... and both proteins are targeted to newly formed cell adhesion sites upon integrin engagement". J. Biol. Chem. 277 (23): 20895- ...
Szasz J, Yaffe MB, Sternlicht H (1993). "Site-directed mutagenesis of alpha-tubulin. Reductive methylation studies of the Lys ...
Biochemical characterization and site-directed mutagenesis". The Journal of Biological Chemistry. 266 (7): 4315-21. doi:10.1016 ...
In vivo site-directed mutagenesis using oligonucleotides. Nat Biotechnol. 19(8):773-6 (Italian words and phrases, Mutagenesis, ... The delitto perfetto technique is also simpler compared to other methods for in vivo site-directed mutagenesis. Other methods ... Delitto perfetto (Italian: [deˈlitto perˈfɛtto]) is a genetic technique for in vivo site-directed mutagenesis in yeast. This ... 2006) The delitto perfetto approach to in vivo site-directed mutagenesis and chromosome rearrangements with synthetic ...
Aldo-keto reductase family 1, member A1
Site-directed mutagenesis of a critical lysine 262". The Journal of Biological Chemistry. 266 (35): 24031-7. doi:10.1016/S0021- ... The tertiary structure consists of a beta/alpha-barrel, with the coenzyme-binding site located at the carboxy-terminus end of ... identification of glycation sites" (PDF). Biochemistry. 34 (4): 1433-8. doi:10.1021/bi00004a038. PMID 7827091. Robinson B, ... characterization of the active site pocket". Biochemistry. 34 (35): 11264-75. doi:10.1021/bi00035a036. PMID 7669785. Takahashi ...
Lounsbury KM, Schlegel B, Poncz M, Brass LF, Manning DR (1993). "Analysis of Gz alpha by site-directed mutagenesis. Sites and ... "Identification of a GTP-binding protein alpha subunit that lacks an apparent ADP-ribosylation site for pertussis toxin". Proc. ...
Another application of site-directed mutagenesis is exchanging an amino acid residue far from the active site with a lysine ... One example of how mutagenesis is used is found in the coupling of site-directed mutagenesis and PCR to reduce interleukin-6 ... Site-directed mutagenesis is a technique that has been around since the 1970s. The early days of research in this field yielded ... Site directed mutagenesis is a valuable technique that allows for the replacement of a single base in an oligonucleotide or ...
This is often investigated through site-directed mutagenesis. In addition, the synthesis of a model complex can suggest the ... Defining the active site structure. A number of important active sites are still poorly defined. This includes the oxygen ... Understanding the active site function. The structure of some enzymes are very well characterized, however, the function of ... ". "Breslow Group Homepage". www.columbia.edu. Retrieved 2015-12-11. Breslow, Ronald (1995-03-01). "Biomimetic Chemistry and ...
2010). "Site-directed mutagenesis of the Anabaena sp. strain PCC 7120 nitrogenase active site to increase photobiological ... Through direct mutagenesis, the scientists exchanged the Asn9 for both Asp (Aspartic Acid) and Glu (Glutamic Acid). This change ... The coenzymes bind to the active site of an enzyme to promote catalysis. By engineering cofactors and coenzymes, a naturally ... The scientists determined that Asn9 (Asparagine, position 9) was an important amino acid the active site of Gre2p. Specifically ...
1999). "Site-directed mutagenesis of restriction endonuclease HindIII". Biosci. Biotechnol. Biochem. 63 (10): 1703-7. doi: ... Despite the uncertainty concerning the structure-catalysis relationship of type II endonucleases, site-directed mutagenesis of ... As a result of the site-mutagenesis experiments previously outlined, it is thus proposed that Lys-125, Asp-123, and Asp-108 of ... Despite the lack of evidence suggesting an exact mechanism for the cleavage of DNA by HindIII, site-mutagenesis analysis ...
Site-directed mutagenesis Restriction digest DNA footprinting Kamvysselis, M. (2003). Computational molecular genomics: genes, ... Non-directed PCR-based mutagenesis can also be used; the parameters of the mutagenic PCR reaction can be adjusted to introduce ... The effects of protein interactions with each other as well as the binding sites can also be assayed in this way; candidate ... This is often necessary because one mutation cannot be guaranteed to inactivate a binding site. ...
... and site-directed mutagenesis of inhibitor-2". The Journal of Biological Chemistry. 269 (2): 944-54. doi:10.1016/S0021-9258(17) ... Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M (November 2006). "Global, in vivo, and site-specific ...
His cloning vectors were also used to develop the method for oligonucleotide site-directed mutagenesis. DNA cloning, shotgun ... sequencing and site-directed mutagenesis became widely used to sequence large DNA molecules like human chromosomes and to ... "Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis". Gene. 26 (1): 101-6. doi:10.1016/0378- ... "Messing's personal website at the Waksman Institute". Waksman.rutgers.edu. Archived from the original on 2010-12-06. Retrieved ...
Marth, J. D. (1996-05-01). "Recent advances in gene mutagenesis by site-directed recombination". The Journal of Clinical ... Ohtsubo, K., and Marth, J.D. (2006). Conditional mutagenesis of the genome using site-specific DNA recombination. In: Gene ... "T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination". Proceedings ... "T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination". Proceedings ...
Chueh PJ, Morré DM, Morré DJ (2002). "A site-directed mutagenesis analysis of tNOX functional domains". Biochim. Biophys. Acta ...
Muscarinic acetylcholine receptor M2
van Koppen CJ, Nathanson NM (December 1990). "Site-directed mutagenesis of the m2 muscarinic acetylcholine receptor. Analysis ...
Site-directed mutagenesis was employed to create mutants in which MgATP binds but does not induce a conformational change. ... Site-specific mutagenesis was used to demonstrate this fact. This has led to a model in which the serine remains coordinated to ... Site-specific mutagenesis has demonstrated that when the lysine is substituted for a glutamine, the protein's affinity for ... Each FeMo cofactor then acts as a site for nitrogen fixation, with N2 binding in the central cavity of the cofactor. The MoFe ...
Stradal TB, Troxler H, Heizmann CW, Gimona M (May 2000). "Mapping the zinc ligands of S100A2 by site-directed mutagenesis". The ... "Transcriptional activation of the tumor suppressor and differentiation gene S100A2 by a novel p63-binding site". Nucleic Acids ... "Repression of the candidate tumor suppressor gene S100A2 in breast cancer is mediated by site-specific hypermethylation". Cell ...
Westheimer et al.'s proposal was further supported by site-directed mutagenesis studies. When Lys116 was mutated to cysteine or ... The active site, consisting of residues such as Phe27, Met97, and Tyr113, is mostly hydrophobic. However, the active site does ... Further research led to the isolation of an active site peptide sequence and identification of the active site lysine, Lys115, ... Arg29 is thought to play a role in substrate binding, while Glu76 is thought to play a role in the orienting the active site ...
Adenine nucleotide translocator
Nelson DR, Lawson JE, Klingenberg M, Douglas MG (April 1993). "Site-directed mutagenesis of the yeast mitochondrial ADP/ATP ... and ADP binding sites of rat liver mitochondria. Soon after, an overwhelming amount of research was done in proving the ...
Multiple cloning sites are sometimes used to ensure that the fragments are inserted in all three possible reading frames so ... Yang WP, Green K, Pinz-Sweeney S, Briones AT, Burton DR, Barbas CF (December 1995). "CDR walking mutagenesis for the affinity ... cDNA has also been analyzed using pIII via a two complementary leucine zippers system, Direct Interaction Rescue or by adding ... Databases, programs and web servers have been widely used to exclude target-unrelated peptides, characterize small molecules- ...
Stabach PR, Cianci CD, Glantz SB, Zhang Z, Morrow JS (1997). "Site-directed mutagenesis of alpha II spectrin at codon 1175 ... Davis LH, Bennett V (1990). "Mapping the binding sites of human erythrocyte ankyrin for the anion exchanger and spectrin". J. ... Bennett AF, Hayes NV, Baines AJ (1991). "Site specificity in the interactions of synapsin 1 with tubulin". Biochem. J. 276. ( ... Steiner JP, Bennett V (1988). "Ankyrin-independent membrane protein-binding sites for brain and erythrocyte spectrin". J. Biol ...
Data has identified VDAC as an interacting partner of BNIP3 and provide direct evidence to support that EndoG is a mediator of ... Wu SL, Li CC, Chen JC, Chen YJ, Lin CT, Ho TY, Hsiang CY (15 January 2009). "Mutagenesis identifies the critical amino acid ... by this gene is a member of the conserved DNA/RNA non-specific ββα-Me-finger nuclease family and possesses a unique site ... During ischemia reperfusion, ROS release substantially contribute to the cell damage and death via a direct effect on the cell ...
DNA damage theory of aging
At most CpG sites cytosine is methylated to form 5-methylcytosine. As indicated in the article CpG site, in mammals, 70% to 80 ... "Nuclear DNA damage as a direct cause of aging" (PDF). Rejuvenation Research. 12 (3): 199-208. CiteSeerX 10.1.1.318.738. doi: ... Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 250 (1-2): 3-16. doi:10.1016/0027-5107(91)90157-j. PMID ... In humans, about 70% of promoters located near the transcription start site of a gene (proximal promoters) contain a CpG island ...
Roger Y. Tsien
... they can be turned fluorescent after deleting some of the signaling parts by genetic means such as site-directed mutagenesis. ... Web page). The Harvard Crimson. Retrieved October 9, 2008. "Cambridge graduate wins Nobel Prize for Chemistry" (Web page). The ... 2004: New "fruit" FPs were generated (by in vitro and in vivo directed evolutions). In 2009, a new kind of Infrared Fluorescent ... One or more of the preceding sentences incorporates text from the royalsociety.org website where: All text published under the ...
A site-directed mutagenesis study". Environ. Health Perspect. 106 (2): 85-92. doi:10.1289/ehp.9810685. PMC 1533021. PMID ... was published suggesting that tropoflavin and various other reported small-molecule TrkB agonists might not actually be direct ...
Site-directed mutagenesis studies have shown that the ultimate length of the isoprenoid product is determined by bulky residues ... Through several more biosynthetic steps, squalene is transformed into lanosterol, a direct precursor for cholesterol. Notably, ... "Bisphosphonates target multiple sites in both cis- and trans-prenyltransferases". Proceedings of the National Academy of ...
"Mutagenesis of conserved residues within the active site of Escherichia coli alkaline phosphatase yields enzymes with increased ... Current researchers are looking into the increase of tumor necrosis factor-α and its direct effect on the expression of ... Alkaline phosphatase contains four Zn ions and two Mg ions, with Zn occupying active sites A and B, and Mg occupying site C, so ... Three metal ions, two Zn and one Mg, are contained in the catalytic sites, and both types are crucial for enzymatic activity to ...
... chemical modification and site-directed mutagenesis of a cephalosporin C acylase from Pseudomonas strain N176". Eur. J. Biochem ...
Howard Ronald Kaback
Kwaw, I; Zen, KC; Hu, Y; Kaback, HR (4 September 2001). "Site-directed sulfhydryl labeling of the lactose permease of ... ISBN 978-3-642-53838-4. Kaback, Howard Ronald; Frillingos S; Sahin-Toth M; Wu J. (1998). "Cys-scanning mutagenesis: a novel ... Kaback, H.R.; Dunten, R.; Frillingos, S.; Venkatesan, P.; Kwaw, I.; Zhang, W.; Ermolova, N. (2007). "Site-directed alkylation ... as well as a battery of site-directed biophysical/biochemical techniques to demonstrate almost incontrovertibly that LacY ...
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing
Too many phosphorothioate bonds can be problematic; each modified site creates a chiral center, which can lead to a racemic ... Ellis, H. M.; Yu, D.; DiTizio, T.; Court, D. L. (2001-06-05). "High efficiency mutagenesis, repair, and engineering of ... Furthermore, the pCas9cr4 plasmid can be purchased from Addgene (ID 62655) for direct implementation into a no-SCAR ... All together, this contributes to a lengthy and inefficient mutagenesis procedure for even single mutations. This technique can ...
"Structure analysis and site-directed mutagenesis of defined key residues and motives for pilus-related sortase C1 in group B ... "Structure analysis and site-directed mutagenesis of defined key residues and motives for pilus-related sortase C1 in group B ... Cleavage occurs between the Thr and Gly, with transient attachment through the Thr residue to the active site Cys residue, ...
... site-directed mutagenesis of active site residues". The Journal of Biological Chemistry. 282 (23): 17157-65. doi:10.1074/jbc. ... One domain facilitates the ATP-grasp mechanism and the other is the catalytic active site for γ-glutamylcysteine. The ATP-grasp ... The domain containing the active site exhibits interesting properties of specificity. In contrast to γ-glutamylcysteine ... Moderately and severely affected individuals have enzymes with dysfunctional catalytic sites, rendering it unable to ...
Raymond C. Stevens
A site-specific mutagenesis, NMR, and X-ray crystallographic study," Prot. Sci. 1, 1435-1446 (1992). Stevens, R. C., & Lipscomb ... all funded by the National Institutes of Health with direct guidance from NIGMS. In 2012, Stevens co-founded the iHuman ... The probing of crystal structure analysis via site-specific mutagenesis," Protein Engineering 4, 391-408 (1991). Gouaux, J. E ... Two disparate ligand binding sites in the human P2Y1 receptor Nature 520: 317-321 Y. Kang, X. E. Zhou, X. Gao, Y. He, W. Liu, A ...
Prostatic acid phosphatase
"Heterologous expression of human prostatic acid phosphatase and site-directed mutagenesis of the enzyme active site". J. Biol. ... 1991). "Covalent structure, disulfide bonding, and identification of reactive surface and active site residues of human ...
Cold Spring Harbor Laboratory
It has been home to eight scientists who have been awarded the Nobel Prize in Physiology or Medicine. CSHL is ranked among the ... See U.S. Patent 2,445,748 (July 27, 1948). Demerec used x-ray mutagenesis to produce a high-yielding strain of Penicillium mold ... During World War Two, Demerec directed efforts at Cold Spring Harbor that resulted in major increases in penicillin production ... Official website Dolan DNA Learning Center Eugenics Archive Cold Spring Harbor Laboratory Press Wikimedia Commons has media ...
DNA polymerase II
One site(s) is responsible for recruiting single-stranded DNA to DNA Pol II and another site(s) is responsible for the ... DNA Polymerase I was the first DNA-Directed DNA polymerase to be isolated from E. coli. Several studies involving this isolated ... Becherel OJ, Fuchs RP (July 2001). "Mechanism of DNA polymerase II-mediated frameshift mutagenesis". Proceedings of the ... The active site contains two Mg2+ ions that are stabilized by catalytic Aspartic Acids D419 and D547. Magnesium ions bind to ...
During repair of DNA double strand breaks, or repair of other DNA damage, incompletely cleared repair sites can cause ... Jacinto FV, Esteller M (July 2007). "Mutator pathways unleashed by epigenetic silencing in human cancer". Mutagenesis. 22 (4): ... homology-directed repair, and DNA methylation". PLOS Genetics. 3 (7): e110. doi:10.1371/journal.pgen.0030110. PMC 1913100. PMID ... "SEER Stat Fact Sheets: All Cancer Sites". National Cancer Institute. Archived from the original on 26 September 2010. Retrieved ...
Most of them occur at CpG sites. DNA methyltransferase is recruited to the site and adds methyl groups to the cytosine of the ... alkylguanine in DNA to itself to fight against mutagenesis and the buildup of toxic compounds that result from alkylating ... which is the overwriting of epigenetic patterns using man-made signals to direct epigenetic proteins to target loci. ... This allows the methyl-CpG binding proteins to bind to the methylated site and cause downregulation of genes. Histone ...
... has a much weaker antiviral effect when its active site has been mutated to the point that the protein can no longer ... Such sublethal mutagenesis contributes to greater genetic diversity among the HIV-1 virus population, demonstrating the ... The predicted deamination reaction is driven by a direct nucleophilic attack on position 4 of the cytidine pyrimidine ring by ... tRNA3Lys typically binds to the HIV-1 primer binding site to initiate reverse transcription. APOBEC3G can inhibit tRNA3Lys ...
... by site-directed mutagenesis". Virology. 187 (2): 654-62. doi:10.1016/0042-6822(92)90468-5. PMID 1546460. Wang MB, Bian XY, Wu ... Parallel to agriculture-directed studies, more basic scientific research elucidated many of viroids' physical, chemical, and ... The enemy at home". RNA Biology. 14 (8): 985-991. doi:10.1080/15476286.2017.1321730. ISSN 1547-6286. PMC 5680766. PMID 28448743 ...
Ball and chain inactivation
... suggesting that TEA competes for an inactivation binding site. Mutagenesis experiments have identified an intracellular string ... Direct inactivation, which occurs in Shaker potassium channels results from the direct blockage of the channel by the ball ... The ball domain enters the channel through the side slits and attaches to a binding site deep in the central cavity. This ... Zhou M, Morais-Cabral JH, Mann S, MacKinnon R (2002). "Potassium channel receptor site for the inactivation gate and quaternary ...
April 1994). "Mutagenesis and mapping of a mouse gene, Clock, essential for circadian behavior". Science. 264 (5159): 719-25. ... Studies also show that Cyclosa turbinata is unique in that its locomotor and web-building activity cause it to have an ... Studies have also helped elucidate how light has a direct effect on human health through its influence on the circadian biology ... February 2001). "An hPer2 phosphorylation site mutation in familial advanced sleep phase syndrome". Science. 291 (5506): 1040-3 ...
Phage-assisted continuous evolution
In these systems, the desired protease cut site is used to link a T7 RNA polymerase and a T7 lysozyme. The T7 lysozyme prevents ... Wang, T.; Badran, A.H.; Huang, T.P.; Liu, D.R. (2018). "Continuous directed evolution of proteins with improved soluble ... Using error-prone polymerases (encoded on the mutagenesis plasmid, or MP), genetic variation is introduced into the protein ... Lane, M.D.; Seelig, B. (2014). "Advances in the directed evolution of proteins". Curr. Opin. Chem. Biol. 22: 129-136. doi: ...
33 (Web Server issue): W690-2. doi:10.1093/nar/gki445. PMC 1160206. PMID 15980564. Chang TH, Huang HD, Wu LC, Yeh CT, Liu BJ, ... Stormo GD, Ji Y (August 2001). "Do mRNAs act as direct sensors of small molecules to control their expression?". Proceedings of ... Unpublished undergraduate research created a similar riboswitch or 'thermosensor' via random mutagenesis of the Listeria ... Abreu-Goodger C, Merino E (July 2005). "RibEx: a web server for locating riboswitches and other conserved bacterial regulatory ...
Wong, T.S.; Tee, K.L.; Hauer, B.; Schwaneberg, U. (2004). "Sequence Saturation Mutagenesis (SeSaM): a novel method for directed ... 2]. SeSaM-Biotech GmbH official homepage. Retrieved April 25, 2017. Official website (Articles needing additional references ... Wong, T.S.; Tee, K.L.; Hauer, B.; Schwaneberg, U. (2004). "Sequence Saturation Mutagenesis (SeSaM): a novel method for directed ... SeSaM-Biotech holds the patent for the Sequence Saturation Mutagenesis method for random mutagenesis, but applies also ...
He is currently professor at École Polytechnique Fédérale de Lausanne (EPFL), where he directs the Laboratory of Cell and ... Pierre Gönczy publications indexed by Google Scholar Publications indexed on ORCID: ORCID 0000-0002-6305-6883 Website of the ... Characterization of Male-Sterile Mutants Generated by Single P Element Mutagenesis". Genetics. 135 (2): 489-505. ISSN 0016-6731 ...
Million RP, Harakawa N, Roumiantsev S, Varticovski L, Van Etten RA (June 2004). "A direct binding site for Grb2 contributes to ... Mutagenesis and Binding assays have helped to identify which molecules and what pathways are downstream of GAB2. The two main ... GAB2 is a large multi-site docking protein (LMD) of about 100kD that has a folded N-terminal domain attached to an extended, ... It is on the C-terminal tail that the various conserved protein binding motifs and phosphorylation sites of GAB2 are found. ...
He received further education in biochemistry at the University of Uppsala 1978 and in site directed mutagenesis in Washington ...
Identification of an important cysteine residue in human glutamate-cysteine ligase catalytic subunit by site-directed...
Molecular Characterization of Binding of Substrates and Inhibitors to DT-Diaphorase: Combined Approach Involving Site-Directed...
Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling. Shiuan Chen, Kebin Wu ... Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling. Shiuan Chen, Kebin Wu ... Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling. Shiuan Chen, Kebin Wu ... Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling ...
How Batrachotoxin Modifies the Sodium Channel Permeation Pathway: Computer Modeling and Site-Directed Mutagenesis - McMaster...
Site-directed Mutagenesis of Ricin A-chain | Soligenix
Site-directed mutagenesis of ricin A-chain and implications for the mechanism of action by Ready, M.P., Y. Kim, and J.D. ... Ready, M.P., Y. Kim, and J.D. Robertus, Site-directed mutagenesis of ricin A-chain and implications for the mechanism of action ... Site-directed mutagenesis of ricin A-chain and implications for the mechanism of action. ...
Ligand binding modes from low resolution GPCR models and mutagenesis: chicken bitter taste receptor as a test-case | Scientific...
... approaches with site-directed mutagenesis was used to characterize the agonist-bound conformation of ggTas2r1 binding site ... Site-directed mutagenesis. In-vitro mutagenesis by PCR-mediated recombination44 was performed as detailed before19, 20, 45 ... For a list of oligonucleotides used for mutagenesis see Table 2.. Table 2 Oligonucleotides used for site-directed mutagenesis ... Residues defining the binding sites were selected for site-directed mutagenesis.. Binding mode predictions of other ggTas2r1 ...
Site Directed Mutagenesis Primer Design
... site directed mutagenesis primer design flexibility by implication to adjacent multiple copies. ... The following the mutagenesis site directed mutagenesis. The direct identification of point mutation site directed mutagenesis ... For mutagenesis site directed mutagenesis kit and care to generate overhanging cohesive ends. Redesign your site directed ... design actually saves a mutagenesis must be two sites and nucleotides. Dna mutagenesis site directed mutagenesis, primer ...
Evidence for OTUD-6B Participation in B Lymphocytes Cell Cycle after Cytokine Stimulation | PLOS ONE
Site-directed Mutagenesis. The OTUD-6B (C188S) mutant was generated by using a QuikChange TM site-directed mutagenesis kit ( ... The putative enhancer region of mouse Otud-6b contains one ETS and one GATA TF binding site. Mutation sites in motifs are ... the functional transcription factor binding sites are very similar as the conserved ETS binding site is required for Otud-6b ... For example, cyclin D2 is a direct target gene of Myc  and PU.1 transcription factor . Its expression can also be ...
Q5® Site-Directed Mutagenesis Kit, New England Biolabs | VWR
... site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hoursThe wide range of reagents are suitable for use ... THE WEB SITE, THE SITE CONTENT, ANY SERVICES PROVIDED ON OR THROUGH THE WEB SITE OR ANY LINKED SITE, INCLUDING ANY DIRECT, ... The Q5® Site-Directed Mutagenesis Kit enables rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 ... including without limitation sites linked to or from the Web Site, sites framed within the Web Site or third party ...
Site-Directed Mutagenesis by Fast & Steep PCR & Seamless Assembly
Steep PCR Site-Directed Mutagenesis and Seamless Assembly @ McGill University. ... needed unique restriction sites withing approximately 1 kb of the mutation sites inorder to perform Site-Directed Mutagenesis. ... Site-Directed Mutagenesis and pEASY Seamless Assembly: *elimination of 1 out of 2 sites of a dual-cut restriction enzyme (Hpa I ... which can be applied to Site-Directed Mutagenesis. We will publish this new protocol in a near future. Combined with Site- ...
Gibson Assembly® Site-Directed Mutagenesis - Tonbo™ A Cytek® Brand
Effects of mutagenesis in the switch I region and conserved arginines of Escherichia coli MnmE protein, a GTPase involved in...
Corporate Fellows | ORNL
Antibody Discovery | IDT
AMOLF Institutional Repository: Molecular Structure of Hydrophobins Studied with Site-Directed Mutagenesis and Vibrational...
Here we use surface-specific sum-frequency generation spectroscopy (VSFG) and site-directed mutagenesis to study the properties ... Molecular Structure of Hydrophobins Studied with Site-Directed Mutagenesis and Vibrational Sum-Frequency Generation ... Molecular Structure of Hydrophobins Studied with Site-Directed Mutagenesis and Vibrational Sum-Frequency Generation ...
Polbase - Reference: Site directed mutagenesis of DNA polymerase I (Klenow) from Escherichia coli. The significance of Arg682...
Site directed mutagenesis of DNA polymerase I (Klenow) from Escherichia coli. The significance of Arg682 in catalysis. ... Site directed mutagenesis of DNA polymerase I (Klenow) from Escherichia coli. The significance of Arg682 in catalysis. Pandey ... we have performed site-directed mutagenesis of this residue. For this purpose the Klenow-coding region of the DNA-pol-I gene ...
Site-directed mutagenesis and PBAN activation of the Helicoverpa zea PBAN-receptor | College of Agricultural Sciences | Oregon...
Site-directed mutagenesis and PBAN activation of the Helicoverpa zea PBAN-receptor ... Site-directed mutagenesis and PBAN activation of the Helicoverpa zea PBAN-receptor ... Site-directed mutagenesis and PBAN activation of the Helicoverpa zea PBAN-receptor. ... For questions or feedback about our college or website, please Contact Us. ...
A Systematic Approach to Defining the microRNA Landscape in Metastasis | Cancer Research | American Association for Cancer...
Site-Directed Mutagenesis of Cytochrome P450scc (CYP11A1). Effect of Lysine Residue Substitution on Its Structural and...
Lepesheva, G. I., Azeva, T. N., Strushkevich, N. V., Gilep, A. A., & Usanov, S. A. (2000). Site-Directed Mutagenesis of ... T1 - Site-Directed Mutagenesis of Cytochrome P450scc (CYP11A1). Effect of Lysine Residue Substitution on Its Structural and ... title = "Site-Directed Mutagenesis of Cytochrome P450scc (CYP11A1). Effect of Lysine Residue Substitution on Its Structural and ... Site-Directed Mutagenesis of Cytochrome P450scc (CYP11A1). Effect of Lysine Residue Substitution on Its Structural and ...
Site directed mutagenesis studies on HIV-1 reverse transcriptase (RT) shed light on the mechanism of action of a new...
Site directed mutagenesis studies on HIV-1 reverse transcriptase (RT) shed light on the mechanism of action of a new ... Site directed mutagenesis studies on HIV-1 reverse transcriptase (RT) shed light on the mechanism of action of a new ... Corona, A., Meleddu, R., Esposito, F. et al. Site directed mutagenesis studies on HIV-1 reverse transcriptase (RT) shed light ... Molecular modeling and site-directed mutagenesis studies supported the hypothesis that RMNC6 could bind to two different RT ...
Specificity of different isoforms of protein phosphatase-2A and protein phosphatase-2C studied using site-directed mutagenesis...
Specificity of different isoforms of protein phosphatase-2A and protein phosphatase-2C studied using site-directed mutagenesis ... Specificity of different isoforms of protein phosphatase-2A and protein phosphatase-2C studied using site-directed mutagenesis ... Specificity of different isoforms of protein phosphatase-2A and protein phosphatase-2C studied using site-directed mutagenesis ... Specificity of different isoforms of protein phosphatase-2A and protein phosphatase-2C studied using site-directed mutagenesis ...
Site-directed mutagenesis and chemical modification of histidine residues on an alpha-class chick liver glutathione S...
Role of Tim17 in coupling the import motor to the translocation channel of the mitochondrial presequence translocase | eLife
Site-directed mutagenesis. Request a detailed protocol Site-directed mutagenesis was done according to standard molecular ... The authors should either provide direct experimental results like crosslinking of BPA in TM1 and TM2 to Tim23 or they should ... suggesting that its binding site should be found in more matrix-facing regions of Tim17. A binding site of Tim44 has not been ... In vivo site-specific crosslinking. Request a detailed protocol Yeast cells expressing BPA-containing versions of Tim17 were ...
2cjy - Proteopedia, life in 3D
Site-directed mutagenesis of PrPc promoter.. Mutation of the p53 putative binding site located on the human PrPc promoter- ... luciferase reporter construct (Mahal et al., 2001) was introduced using the Quick change site-directed mutagenesis kit ( ... 6G, compare WT/p53 and Mut/p53). This set of data brings the demonstration that p53 could act as a direct activator of PrPc ... It should be noted, however, that the PRNP gene did not emerge when a global mapping of p53-binding sites in the human genome ...
Position Avaialble, Molecular Microbial Ecology
Onur Dagliyan, Ph.D. | Harvard Catalyst Profiles | Harvard Catalyst
View source - 2009.igem.org
Site-Directed Mutagenesis ,/font>,/b>,/i> ,br> ,p>QuikChange II Site-Directed Mutagenesis Kit (Stratagene) ,/p> ,ol> ,li> ... each reaction using the cycling parameters as outlined in Table I of the Stratagene QuikChange II Site-Directed Mutagenesis Kit ... Introduction also to the 2009 iGEM home page and our team wiki. ,/p> ,p class="header">June 23,/p> ,p class = "class">After ... HOME,/font>,/a>,/td> ,/tr> ,tr> ,td id="nav">,a href="https://2009.igem.org/Team:Utah_State/Team">,font size = 4>ABOUT US,/font ...
Yuzhu Zhang : USDA ARS
Thermostability enhancement of cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus by site-directed mutagenesis - ... Thermostability enhancement of cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus by site-directed mutagenesis. ... Official websites use .gov A .gov website belongs to an official government organization in the United States. ... ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Healthy Processed Foods Research » ...
SMART: ANK domain annotation
Site-directed mutagenesis of these residues confirms their importance in ARF-GAP activity. ... However, the role of residues 348-568 may be at the level of folding rather than a direct contact since the deleted sequences ... An invariant arginine and several nearby hydrophobic residues are solvent exposed and are predicted to be the site of ... Structure of Gankyrin-S6ATPase photo-cross-linked site-specifically, and incoporated by genetic code expansion. ...
ResiduesPolymeraseOligonucleotide-DirectedVariantsAminoProteinSubunitResidueMutationMutantsMutantVitroComputer ModelingExperimentsCloningEfficientEnzymesUnique restrictionProteinsOligomericSynthesisInteractionTransgeneHumansRestriction enzymeGeneProteaseMotifCleavageBiochemistryConsistsDemonstratePathwayLiverRodentGenomeStudySubstitutionPhosphorylationAdjacentSingleRegionBinding SitesProcessSequenceStructureProductMechanismCookiesDetailsLargeStudiesWorkProductionSpecific sites
- To identify cysteine residues in GLCL that are involved in its activity, eight conserved cysteine residues in human GLCL catalytic subunit (hGLCLC) were replaced with glycine residues by PCR-based site-directed mutagenesis. (portlandpress.com)
- It does not involve multiple site mutagenesis has attracted funding from those residues in. (west-covina-lawyer.com)
- The present study was undertaken to further evaluate the role of these residues in the interaction of cytochrome P450scc with adrenodoxin using site-directed mutagenesis. (skoltech.ru)
- Residues hypothesized to be critical for compound binding were mutated by site-directed mutagenesis. (biomedcentral.com)
- Mutagenesis results identified few amino acid residues in the RNase H domain that are essential for RNase H inhibition by RMNC6 but do not influence its DNA polymerase inhibition. (biomedcentral.com)
- Site-directed mutagenesis and chemical modification of histidine residues on an alpha-class chick liver glutathione S-transferase CL 3-3. (sinica.edu.tw)
- An invariant arginine and several nearby hydrophobic residues are solvent exposed and are predicted to be the site of interaction with ARFs. (embl.de)
- Site-directed mutagenesis of these residues confirms their importance in ARF-GAP activity. (embl.de)
- Protein sequence alignment, structure modeling and site-directed mutagenesis demonstrated that Ser 10 , Gly 48 , Asn 77 , Asp 158 and His 161 residues composed the active centre of 'AcTesA. (biomedcentral.com)
- Further studies using the yeast two-hybrid system revealed that the NS5 region (residues 320-368) immediately adjacent to the NLS contained an importin - binding site that abuts or overlaps the binding site for the NS3 protein (protease/helicase). (who.int)
- To determine which amino acid residues are essential for the catalytic activity of mouse Gal1,3GalNAc 2,3-sialyltransferase (mST3Gal I), chemical modification and site-directed mutagenesis were employed against tryptophan and cysteine residues located in the predicted catalytic domain. (who.int)
- Since this lysozyme contained two amino acid substitutions (Ala 31 →Valand Asn 106 →Ser)in addition to an extra methionine residue at the NH 2 -terminus the substituted amino acid residues were converted back to the original ones by means of oligonucleotide-directed site-specific mutagenesis and in vitro recombination. (elsevier.com)
- Reference: Site directed mutagenesis of DNA polymerase I (Klenow) from Escherichia coli. (neb.com)
- Blind docking analysis suggested that RMNC6 could bind to two different RT pockets: the first one close to the RNase H active site, the second one near to the polymerase active site. (biomedcentral.com)
- Binding to the RNase H site appears to be responsible for RNase H inhibition, while binding to the DNA polymerase site seems to affect both RT-associated functions by both short- and long-range effects. (biomedcentral.com)
- Oligonucleotide-directed mutagenesis was used to construct chimeric cDNAs that encode the extracellular and transmembrane domains of the vesicular stomatitis virus glycoprotein (G) linked to the cytoplasmic domain of either the immunoglobulin mu membrane heavy chain, the hemagglutinin glycoprotein of influenza virus, or the small glycoprotein (p23) of infectious bronchitis virus. (rupress.org)
- Researchers can then create variants via mutagenesis or by using synthetic dsDNA through an iterative process to further screen and select which molecule to move forward in the workflow. (idtdna.com)
- This dataset corresponds to a comprehensive splicing analysis of splice-site variants of exons 1 to 3 of the breast cancer susceptibility gene PALB2. (fecyt.es)
- Three-step site-directed mutagenesis screen identifies pathogenic MLH1 variants associated with Lynch syndrome. (cdc.gov)
- Lfeap mutagenesis experiment if single pcr protocols were used in directed mutagenesis pcr fragment of amino acid transporters. (west-covina-lawyer.com)
- The present results, together with previous findings, suggest that the changes in functional activity of mutant Lys405Gln may reflect the direct participation of this amino acid residue in the electrostatic interaction of cytochrome P450scc with its physiological partner, adrenodoxin. (skoltech.ru)
- Plant cystatins, similar to other defense proteins, include hypervariable, positively selected amino acid sites presumably impacting their biological activity. (usda.gov)
- Three primer design primers designed in directed protein. (west-covina-lawyer.com)
- For advances in protein structure and enzyme mechanisms by use of affinity labeling and site-directed mutagenesis. (ornl.gov)
- We have expressed the catalytic domain of Chinese hamster HMG-CoA reductase, and 13 point mutations involving the region around the single phosphorylation site for AMP-activated protein kinase. (elsevier.com)
- By fusing the jellyfish enhanced green fluorescent protein reporter molecule (EGFP) to the carboxy-terminus of Us9, we demonstrated that Us9 not only is capable of targeting a Us9-EGFP fusion protein to the Golgi compartment, it also is able to direct efficient incorporation of such chimeric molecules into infectious viral particles. (vetres.org)
- Custom services include but are not limited to RNA and cDNA preparation, cloning, site-directed mutagenesis, protein expression, and custom PCR assays. (harvard.edu)
- Students in the course now design their own mutant of γD crystallin (a human protein implicated in congenital and age-onset cataractogenesis ) using site-directed mutagenesis, purify and express their protein, and then study its stability using fluorescence and AFM. (brandeis.edu)
- Site directed mutagenesis experiments showed that reducing the positive charge of this surface-exposed region of the E protein DII correlated with improved Ae. (cdc.gov)
- Bolded and underlined sequences were modeled with poly-alanine structural elements, and the large subunit tRNA binding sites, providing a reversible ribosome inactivation mechanism. (kellersign.com)
- In order to further define the role of Arg682 in the catalytic process, we have performed site-directed mutagenesis of this residue. (neb.com)
- Site-directed mutagenesis of Asp179 confirmed the significance of this residue in substrate recognition. (proteopedia.org)
- To introduce the desired stop codons without performing PCR amplification of the whole pGOI vector, we needed unique restriction sites withing approximately 1 kb of the mutation sites inorder to perform Site-Directed Mutagenesis. (civicbio.com)
- Our Mutagenesis Portfolio offers several mutation and library services to meet our various customer needs. (genscript.com)
- A mutation affecting the stability of the stem-loop structure immediately downstream of the processing site had two effects. (microbiologyresearch.org)
- design and creation of Stop codon mutants by Fast & Steep PCR* Site-Directed Mutagenesis and pEASY Seamless Assembly or restriction enzyme/T4 ligase cloning. (civicbio.com)
- Growth is designed primers and mutagenesis site directed mutagenesis reaction pathways that mutant primer designing primers were performed larger plasmids explained in joining sites. (west-covina-lawyer.com)
- Further structure-guided mutagenesis generated a quadruple mutant (Y50F/N110S/I212T/L264V), whose K_M for isobutyraldehyde is ∼17-fold lower and catalytic efficiency (k_(cat)/K_M) is ∼160-fold higher than wild-type LlAdhA. (caltech.edu)
- Since the wild type enzyme (Met -1 -lysozyme) was activated in vitro without loss of specific activity, the systems described in this study (mutagenesis, overproduction, purification and folding of inactive mutant lysozymes) may be useful in the study of folding pathways, expression of biological activity and stability of lysozyme. (elsevier.com)
- 80 (Pt 2), 291-296 (1999) PUBMED 10073687 REFERENCE 3 (bases 1 to 7654) AUTHORS Liu,B., Clarke,I.N. and Lambden,P.R. TITLE Polyprotein processing in Southampton virus: identification of 3C-like protease cleavage sites by in vitro mutagenesis JOURNAL J. Virol. (cdc.gov)
- Recent developments in site-directed mutagenesis, computer-modeling, and mechanistic analysis of cytochromes P450 and flavin-containing monooxygenases are described. (metu.edu.tr)
- Site-directed mutagenesis experiments demonstrate that tyrosines 600 and 778 of EphB1 are required for its interaction with c-Src and p52 Shc . (rupress.org)
- Trees displayed for a single mutagenic sites in this fact avoids cloning procedures as you could be evaluated from almost dna sequencing reaction efficiency. (west-covina-lawyer.com)
- Combined with Site-Directed Mutagenesis and pEASY Cloning by Seamless Assembly , this protocol also allows for multisite-directed mutagenesis . (civicbio.com)
- Adenoviral vectors, particularly human serotype 5 (of 58 individual serotypes), are widely used for gene therapy due to versatile tropism, large transgene capacity and ability to achieve efficient transgene without risk of insertional mutagenesis. (ukri.org)
- The nickel ion, the mid these strategies that primer vendor of bases in directed mutagenesis site in a unique restriction enzymes to adjacent sequences. (west-covina-lawyer.com)
- We explored the enzymes responsible for directing UT-A3 sialylation. (emory.edu)
- The Original WT Plasmid Map does not have Unique Restriction Enzyme sites anywhere near the desired Stop Codon Insertion sites. (civicbio.com)
- Site-directed mutagenesis was used to selectively eliminate the IgG binding for each epitope and single and multiple mutations were expressed as recombinant GST fusion proteins. (cdc.gov)
- One way is to stabilize one particular oligomeric form using site-directed mutagenesis. (news-medical.net)
- As a direct synthesis and subsequent annealing that can simplify routine mutagenesis? (west-covina-lawyer.com)
- Interestingly, the coated virus then only delivers genes to the liver via the interaction with the coagulation factor and not a direct interaction between the liver and the viru (i.e. the coagulation factor acts as a bridge between the cell and the virus). (ukri.org)
- The most widely used transgenesis method utilizes the yeast meganucleases I-SceI which cuts both the transgene vector and an unknown site in the genome into which the transgene gets integrated. (brandeis.edu)
- The custom meganucleases engineered by Cellectis Bioresearch target a known single site (24bp) within the genome, allowing for increased specificity and efficiency of transgene intergration. (brandeis.edu)
- In this chapter, electrophilic agents nogenicity from studies of exposed (benzene, 1,3-butadiene, and eth- include direct-acting electrophilic humans. (who.int)
- Public health officials and others concerned with appropriate actions to take at hazardous waste sites may want information on levels of exposure associated with more subtle effects in humans or animals (LOAEL) or exposure levels below which no adverse effects (NOAELs) have been observed. (cdc.gov)
- Elimination of 1 out of 2 sites of a dual-cut restriction enzyme (Hpa I) and addition of a unique cutter (Xho I) and addition of a Kpn I site next to the stop codon. (civicbio.com)
- To protect the research and privacy of the customer who paid for our Design, Single Site- and MultiSite-Directed Mutagenesis Services , the name of the gene of interest and researcher are held confidential. (civicbio.com)
- Seven polymorphic sites in the beta-globin gene cluster were analyzed on a sample of 96 chromosomes of Venezuelan sickle cell patients from the State of Aragua. (embrapa.br)
- 78 (9), 4827-4837 (2004) PUBMED 15078964 REFERENCE 2 (bases 1 to 7654) AUTHORS Liu,B.L., Viljoen,G.J., Clarke,I.N. and Lambden,P.R. TITLE Identification of further proteolytic cleavage sites in the Southampton calicivirus polyprotein by expression of the viral protease in E. coli JOURNAL J. Gen. Virol. (cdc.gov)
- The acidic motif, however, does not contain signals needed to direct the incorporation of Us9 into viral envelopes. (vetres.org)
- Site-directed mutagenesis of the dileucine motif results in an increase in Us9 plasma membrane staining and a partial internalisation defect. (vetres.org)
- Mutations were introduced in the region of the cleavage site. (microbiologyresearch.org)
- For more information, please visit the Department of Chemistry and Biochemistry website . (laurentian.ca)
- At Civic Bio, we rarely use a site-directed mutagenesis strategy whcih consists in amplifying the whole vector (read PCR Success Story #16 for the difference between 2 workflows). (civicbio.com)
- The C5b6 interface consists of three binding sites stabilized predominantly by van der Waals interactions, and several critical salt bridges and hydrogen bonds. (frontiersin.org)
- We demonstrate a widely applicable solution: targeted genome editing using site-specific nucleases. (bioseek.eu)
- Our study is the first demonstration of the involvement of presenilins in the control of PrP c expression and reveals a new pathway linking γ-secretase and p53 to the direct control of PrP c transcription. (jneurosci.org)
- UDS activity in the rat liver of the human carcinogens benzidine and 4-aminobiphenyl, and the rodent carcinogens 3,3'-dichlorobenzidine and direct black 38. (cdc.gov)
- We conclude that at least three ADV genes (gE, gI and Us9) are all necessary, but each is not sufficient to sponsor anterograde directed spread of the virus in specific rodent neurons. (vetres.org)
- Meganucleases are endodeoxyribonucleases characterized by a large recognition site (12 to 40 base pairs) - so large that it generally only occurs once in any given genome. (brandeis.edu)
- Here we use surface-specific sum-frequency generation spectroscopy (VSFG) and site-directed mutagenesis to study the properties of class I hydrophobin (HFBI) films from Trichoderma reesei at the molecular level. (amolf.nl)
- Sequential methionine substitution by SPOTsalogue identified K68, E122, and K131 as critical aa in each epitope to change by site-directed mutagenesis. (cdc.gov)
- Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion. (bvsalud.org)
- Deletion analysis of the cytoplasmic tail reveals that an acidic cluster containing putative phosphorylation sites is necessary for the recycling of Us9 from the plasma membrane. (vetres.org)
- introduction of a unique cutting site (Kpn I) adjacent to the original Stop codon. (civicbio.com)
- We conclude that each one of these promoter regions has a particular arrangement, either a single CtrA binding site for activation of fliL2p and fliF2p , or two independent sites for activation of flgB2p and fliI2p . (biomedcentral.com)
- A site-directed mutagenesis verified a single glycosylation site in UT-A3 at Asn279. (emory.edu)
- Binding sites for CtrA of these promoters were identified in silico and tested by site directed mutagenesis. (biomedcentral.com)
- We analyzed these regions and found that the arrangement of the CtrA binding sites varies considerably. (biomedcentral.com)
- Mutagenesis process and mutagenesis at methylated and then test for directed mutagenesis libraries ranging from roche. (west-covina-lawyer.com)
- Because of low sequence similarity between ggTas2r1 and crystallized GPCRs (~10% identity, ~30% similarity at most), the combination of computational approaches with site-directed mutagenesis was used to characterize the agonist-bound conformation of ggTas2r1 binding site between TMs 3, 5, 6 and 7. (nature.com)
- Maltose binding site 2 mutations affect product inhibition of Bacilluscirculans stb01 cyclodextrin glycosyltransferase. (usda.gov)
- Ready, M.P., Y. Kim, and J.D. Robertus, Site-directed mutagenesis of ricin A-chain and implications for the mechanism of action . (soligenix.com)
- These cookies allow us to count visits and traffic sources so we can measure and improve the performance of our site. (cdc.gov)
- If you do not allow these cookies we will not know when you have visited our site, and will not be able to monitor its performance. (cdc.gov)
- Cookies used to make website functionality more relevant to you. (cdc.gov)
- Schematic details of the generation of multiple mutations with LFEAP mutagenesis. (west-covina-lawyer.com)
- A large number of publications and patents based upon mutagenesis studies support this correlation (see e.g. (justia.com)
- This method has several limitations: it requires a large number of embryos for injection and screening, the integration sites cut by I-Sce-I are unknown and likely stochastic, and it ultimately produces only 5-10% of germline transmission. (brandeis.edu)
- Molecular modeling and site-directed mutagenesis studies supported the hypothesis that RMNC6 could bind to two different RT sites. (biomedcentral.com)
- Our website will not work properly. (rcsb.org)
- Combining detailed structural information and directed evolution, we have achieved significant improvements in non-native alcohol dehydrogenase activity that will facilitate the production of next-generation fuels such as isobutanol from renewable resources. (caltech.edu)
- The researcher located at McGill University in Montreal contacted us to introduce Stop codons at specific sites using Site-Directed Mutagenesis . (civicbio.com)
- Glucosyltransferases synthesize exopolysaccharides, which are glucans that promote the accumulation of microorganisms at specific sites on dental surfaces, and glucosyltransferases become enzymatically active when exposed to dietary sucrose. (bvsalud.org)