Principle: The familiar "BioBrick cloning" enzymes (i.e., EcoRI, NotI, XbaI, SpeI, PstI) are Type II restriction enzymes, which cut the sequences that they specifically bind to. The Type IIS Assembly method uses a Type IIS restriction enzyme, which binds at a specific sequence and cuts at a non-specific location exactly five base pairs away. As a result, the enzyme cleaves away its own binding site and leaves behind the most useful feature of assembly, sticky overhangs. When designed properly, Type IIS sites can be used to perform seamless assembly of parts. As an added convenience, this protocol allows cutting and ligation to occur in a single tube, as a single reaction. Thus, gel purification steps can be eliminated. This protocol uses the Type IIS restriction enzyme BsmBI (CGTCTCn/nnnn). ...
Principle: The familiar "BioBrick cloning" enzymes (i.e., EcoRI, NotI, XbaI, SpeI, PstI) are Type II restriction enzymes, which cut the sequences that they specifically bind to. The Type IIS Assembly method uses a Type IIS restriction enzyme, which binds at a specific sequence and cuts at a non-specific location exactly five base pairs away. As a result, the enzyme cleaves away its own binding site and leaves behind the most useful feature of assembly, sticky overhangs. When designed properly, Type IIS sites can be used to perform seamless assembly of parts. As an added convenience, this protocol allows cutting and ligation to occur in a single tube, as a single reaction. Thus, gel purification steps can be eliminated. This protocol uses the Type IIS restriction enzyme BsmBI (CGTCTCn/nnnn). ...
294449241 - EP 1054959 A1 2000-11-29 - HIGH EFFICIENCY METHODS AND COMPOSITIONS FOR INTEGRATING EXOGENOUS DNA INTO GENOMIC DNA OF SPERM - [origin: WO9942569A1] The present invention relates to high efficiency methods and compositions for stably integrating exogenous double-stranded DNA into the chromosomal DNA of sperm cells. The exogenous DNA to be integrated into the sperm is converted to a linear double stranded DNA possessing single-stranded cohesive ends, by contacting said exogenous DNA with a type II restriction enzyme that upon scission, generates such ends. The DNA to be cut can be a circular DNA such as in a plasmid, such that it possesses at least one recognition and cutting site outside of the genes or regulatory regions critical to the desired, post-integration function of the DNA, and no recognition and cutting sites within the critical regions. The exogenous DNA is then introduced into sperm cells, as is some of the type II restriction enzyme used above to generate the single-stranded
Type II restriction enzymes are the molecular scissors that catalyse the double‐strand cleavage of deoxyribonucleic acid (DNA) at specific base sequences
Plasmid ColIb-P9 (Incll) encodes mechanisms which allow it to avoid destruction by type I and type II restriction enzymes during transfer by conjugation between strains of Escherichia coli. A genetic system was developed to analyse these mechanisms. The system relied on measuring Collb-mediated rescue of the restriction-sensitive plasmid R751 (IncPp) from destruction by EcoKI (type I) and EcoRI (type II). One Collb mechanism was known to involve a plasmid-encoded antirestriction gene known as ardA, the product of which is active against type I enzymes. Tests for alleviation of EcoKI restriction of R751, showed strong protection by a co-transferring Collb (Ard+) plasmid, slight protection when Collb was resident in the recipient and no effect when Collb was immobilised in the donor by removal of its nic site. Hence, expression of ardA is activated in the recipient cell following transfer no detectable transfer of the ArdA protein occurs from the donor to the recipient. The ardA gene is found in ...
endodeoxyribonuclease AclI: type II restriction enzyme, EC 3.1.21.4, from Acinetobacter calcoaceticus; recognizes 5-AA/CGTT-3 and cleaves at the slash
FUNCTION: Methylates DNA within the sequence GATC and protects the DNA from cleavage by the restriction endonuclease MboI. Although it shares sequence specificity with a number of type II restriction endonucleases and methylases, it is thought to act in postreplication mismatch repair rather than as a part of a restriction modification system. May also play a role in DNA replication ...
A new Type II restriction endonucleaseApaCI purified fromAcetobacter pasteurianus is an isoschizomer ofBamHI that cleaves at the nucleotide sequence 5′-G/GATCC-3′ of double-stranded DNA. The single restriction activity present in this strain permits rapidly purified 30 000 units of cleavage activity from 10 g of freshly harvested cells. The resultingApaCI preparation is free of contaminant nuclease activities that might interfere within vitro manipulation of DNA.
We believe that new standards should only be introduced to the Registry of Standard Biological Parts if they are compatible with the existing standards (most importantly with RFC10). Otherwise, the registry would get functionally split up into smaller part libraries and teams using one standard could not collaborate with teams using another. We therefore were very careful with introducing type IIs restriction sites into iGEM backbones. In this context, choosing the optimal relative position of the type IIs binding site to the BioBrick prefix and suffix restriction sites is essential for preserving idempotency of the RFC 10 standard. Idempotency in this context means that assembling BioBricks results in higher order constructs that meet BioBrick standard requirements (i.e. they are flanked by the four standard restriction sites and do not contain any of them). The type IIs restriction site can principally be placed in three different positions ...
This is an interface to a utility that reports all intervals of a desired size between cutting sites of either a single restriction enzyme, or a pair of enzymes. All cutting sites are determined in the human sequence. Error tolerance for interval sizes is selectable, as well as the direction of matching for the determination of enzyme cutting sites. First enzyme (mandatory): AatII AccI Acc65I AciI AflII AflIII AgeI AluI AlwI AlwNI ApaI ApaLI ApoI AscI AseI AvaI AvaII AvrII BamHI BanI BanII BbsI BbvI BcgI BcgI(second_site) BclI BfaI BglI BglII BpmI Bpu1102I BsaI BsaAI BsaBI BsaHI BsaJI BsaWI BsgI BsiEI BsiHKAI BsiWI BslI BsmI BsmAI BsmFI Bsp120I Bsp1286I BspDI BspEI BspHI BspMI BsrI BsrBI BsrDI BsrFI BsrGI BssHII Bst1107I BstBI BstEII BstNI BstUI BstXI BstYI Bsu36I ClaI Csp6I DdeI DpnI DpnII DraI DraIII DrdI EaeI EagI Eam1105I EarI Ecl136II Eco47III Eco57I EcoNI EcoO109I EcoRI EcoRV Esp3I Fnu4HI FokI FspI HaeII HaeIII HgaI HhaI HincII HindIII HinfI HinP1I HpaI HpaII HphI KasI KpnI MboI MboII MluI ...
This is an interface to a utility that determines the locations at which the selected enzyme is cutting the human sequence. If so specified, alternative enzymes that cut the sequence at the same location are reported. Enzyme: AatII AccI Acc65I AciI AflII AflIII AgeI AluI AlwI AlwNI ApaI ApaLI ApoI AscI AseI AvaI AvaII AvrII BamHI BanI BanII BbsI BbvI BcgI BcgI(second_site) BclI BfaI BglI BglII BpmI Bpu1102I BsaI BsaAI BsaBI BsaHI BsaJI BsaWI BsgI BsiEI BsiHKAI BsiWI BslI BsmI BsmAI BsmFI Bsp120I Bsp1286I BspDI BspEI BspHI BspMI BsrI BsrBI BsrDI BsrFI BsrGI BssHII Bst1107I BstBI BstEII BstNI BstUI BstXI BstYI Bsu36I ClaI Csp6I DdeI DpnI DpnII DraI DraIII DrdI EaeI EagI Eam1105I EarI Ecl136II Eco47III Eco57I EcoNI EcoO109I EcoRI EcoRV Esp3I Fnu4HI FokI FspI HaeII HaeIII HgaI HhaI HincII HindIII HinfI HinP1I HpaI HpaII HphI KasI KpnI MboI MboII MluI MnlI MscI MseI MslI MspI MspA1I MunI MwoI NaeI NarI NciI NcoI NdeI NgoMI NheI NlaIII NlaIV NotI NruI NsiI PacI PaeR7I PflMI PleI PmeI PmlI Ppu10I PpuMI ...
dszA has four PstI cut sites. dszB has a PstI and a NotI cut site. dszC has two PstI cut sites. In order to be able to use these parts in construction, we had to eliminate these cut sites. Stratagene site directed mutagenesis was used for this purpose. During this procedure, two primers are designed for each mutation. Primers are complementary to the DNA strands except for one base pair. Therefore after the PCR, the whole plasmid is amplified with a point mutation. The base pair was chosen in a way so that it eliminates the cut site but the codon it belongs to still codes for the same amino acid (a silent mutation). We also used the same method to perform the Y63F mutagenesis in dszB (rationale will be explained). In this case the primers also introduce a point mutation. The point mutation causes the codon to code for Phe instead of Tyr. We also did this in a way so that the site of the mutation turns into a hpyAV cut site so we can confirm the mutagenesis by cutting the plamid with ...
Restriction and modification systems are wide spread among prokaryotes and pre-sent an efficient protection against invasion of mobile genetic elements. In general, they code for a restriction endonuclease (REase) and a DNA-methyltransferase (MTase) of the same DNA specificity. The MTase methylates the cellular DNA and by this epigenetic marker protects it against the action of the REase. The REase pre-vents the entry of foreign unmethylated DNA by site-specific cleavage. EcoRII is an REase which needs at least two copies of the recognition sequence for efficient cleavage. Investigations of the EcoRII structure and function revealed that the pro-tein is composed of two stable domains: the N-terminal domain acts as a repressor by sterically blocking the C-terminal domain and thereby inhibiting its catalytic activity. This regulatory mechanism is known as autoinhibition and has been often described for eukaryotic proteins, but for the first time was proposed for a REase. In this work, we verified ...
Any recognition sequence that was four nucleotides in length could be found every 256 nucleotides (on the average) in this simple scenario. In actuality, sequences are usually not evenly made up of G, A, T, and C nucleotides, which skews the statistics a bit. In addition, certain short sequences may be more or less common in the DNA, which will also affect the frequency with which a recognition sequence is found. The dinucleotide CG is very uncommon in mammalian DNA, which makes it less likely that you will find a recognition sequence for the enzyme Hpa II (C^CGG).. Longer recognition sequences lead to lower probability of having a site at any point in a DNA strand. In our simplistic scenario where every nucleotide is evenly distributed in DNA, you would expect to find a PacI site every 65,000 nucleotides (on the average). Thats because theres a one in four chance that each of the eight nucleotides (taken individually) in a random sequence is just right for PacI. The chances of being lucky ...
The PDB archive contains information about experimentally-determined structures of proteins, nucleic acids, and complex assemblies. As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.
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 ...
Generation of transgenic mice. An 8.5 kb upstream region of the CaMKIIα gene was replaced with cytomegalovirus (CMV) promoter of pCMVβ vector (Clontech, Cambridge, UK), and the rat cDNA of NMDAR2D was subcloned into the NotI site of this plasmid. The insert was purified by sucrose density gradient and injected into the pronucleus of fertilized eggs. Hybrids of C57Bl/6 and C3H (B6C3) were used as hosts of transgene and were backcrossed six to seven times onto C57Bl/6 background. Lines 9 and 17 were used primarily in this study. Basically identical results were obtained with these two lines, and data from line 9 were presented except where noted. The genetic contribution of C57Bl/6 and C3H strains at the generation when the transgenic mice were analyzed was 1.6 and 98.4% for line 9, respectively, and 0.8 and 99.2% for line 17, respectively. Control wild-type mice were chosen from the same littermates. For the electrophysiology of adult hippocampus and behavioral experiments, the same groups of ...
Database UniCarb-DB. Taxonomical restrictions None. Other restrictions None. Allowed cleavages None. Parent error 0.5 m/z. Fragment error 0.5 m/z. Mass accuracy Scoring method Spectral matching. Scoring algorithm dot product. Scoring result 0.99,Identical,1. Scoring value format UniCarb-DB triplet. ...
Database UniCarb-DB. Taxonomical restrictions None. Other restrictions None. Allowed cleavages None. Parent error 0.5 m/z. Fragment error 0.5 m/z. Mass accuracy Scoring method Spectral matching. Scoring algorithm dot product. Scoring result 0.99,Identical,1. Scoring value format UniCarb-DB triplet. ...
My baby is 6 months old. Obviously I realize she is not weaning, because I think that would be against her biology, since she has never had a bottle or paci or anything but me. Heres the trouble, she has become less and less interested in nursing. In fact, now she rolls away from me most of the time when I offer to feed her. Her output is fine, 3 poops/day. I have her 6 month appt next week so Ill know about her weight then but so far its been the high end of normal. She only eats when
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... - Reactie specifica de aparare a organismului, caracterizat prin adunarea in mici gramezi a globulelor rosii, bacteriilor sau altor elemente, in prezenta anticorpilor corespunzatori
Hamilton Othanel Smith (1931 - present) was awarded one-third of the Nobel Prize for Physiology or Medicine in 1978 for his discovery of a class of restriction enzymes (used by the cell to splice two pieces of DNA) that recognize specific sequences of nucleotides in DNA and cleave it at particular points. While studying how the bacterium Haemophilus influenzae takes up DNA from the phage virus P22, Smith and his colleagues discovered the first of what came to be called type II restriction enzymes. These enzymes not only recognize a specific region in a DNA sequence but always cut the DNA at that very site. This predictable behavior made type II restriction enzymes valuable tools in the study of DNA structure and in recombinant DNA technology (in which artificial DNA is made by combining other DNA sequences together).. ...
Define restriction endonuclease. restriction endonuclease synonyms, restriction endonuclease pronunciation, restriction endonuclease translation, English dictionary definition of restriction endonuclease. Noun 1. restriction endonuclease - any of the enzymes that cut nucleic acid at specific restriction sites and produce restriction fragments; obtained from...
Author: Grazulis, S. et al.; Genre: Journal Article; Published in Print: 2002-02-15; Title: Crystal structure of the Bse634I restriction endonuclease: comparison of two enzymes recognizing the same DNA sequence
Zhu, M.; Yu, J.; Zhou, C.; Fang, H., 2016: Markerless DNA deletion based on Red recombination and in vivo I-Sec I endonuclease cleavage in Escherichia coli chromosome
This article contains a list of the most studied restriction enzymes whose names start with Ba to Bc inclusive. It contains approximately 120 enzymes. The following information is given: Enzyme: Accepted name of the molecule, according to the internationally adopted nomenclature, and bibliographical references. (Further reading: see the section "Nomenclature" in the article "Restriction enzyme".) PDB code: Code used to identify the structure of a protein in the PDB database of protein structures. The 3D atomic structure of a protein provides highly valuable information to understand the intimate details of its mechanism of action. Source: Organism that naturally produces the enzyme. Recognition sequence: Sequence of DNA recognized by the enzyme and to which it specifically binds. Cut: Cutting site and DNA products of the cut. The recognition sequence and the cutting site usually match, but sometimes the cutting site can be dozens of nucleotides away from the recognition site. Isoschizomers and ...
XbaI and PvuII Polymorphisms of Estrogen Receptor 1 Gene in Females with Idiopathic Scoliosis: No Association with Occurrence or Clinical Form. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
pVTR. The pVTR vector series is designed to be used for gene expression under lac-inducer systems, such as IPTG. This method is based on VTR expression cassettes, which consist of a 1.98 Kb sequence containing the gen lacIq next to the Ptro promotor and the laco operator for lacIq repressor. The cassette also contains an optimized TIR which includes a start codón ATG overlapping with the restriction site NcoI.. The three cassettes are flanked by NotI targets and are available in low copy number plasmids (pVTR-A, pVTR-B and pVTR-C) with pSC101 ori. They can be selected for resistance to chloramphenicol.. pVTR vectors are useful for gene conditional expression prior to transference, (as NotI restriction fragments), to the NotI site of any pUT-miniTn5 vectors for the final integration into the chromosome. ...
Standardized DNA assembly strategies facilitate the generation of multigene constructs from collections of building blocks in plant synthetic biology. A common syntax for hierarchical DNA assembly following the Golden Gate principle employing Type IIs restriction endonucleases was recently developed, and underlies the Modular Cloning and GoldenBraid systems. In these systems, transcriptional units and/or multigene constructs are assembled from libraries of standardized building blocks, also referred to as phytobricks, in several hierarchical levels and by iterative Golden Gate reactions. This combinatorial assembly strategy meets the increasingly complex demands in biotechnology and bioengineering, and also represents a cost-efficient and versatile alternative to previous molecular cloning techniques. For Modular Cloning, a collection of commonly used Plant Parts was previously released together with the Modular Cloning toolkit itself, which largely facilitated the adoption of this cloning ...
DNA Restriction Enzymes from Takara such as Sau3AI are high-quality: perform restriction enzyme digestion with reliable restriction endonucleases
Restriction enzymes are also called molecular scissors as they cleave DNA at or near specific recognition sequences known as restriction sites. These enzymes make one incision on each of the two strands of DNA and are also called restriction endonucleases.4, 5. Viruses infect the host cells by injecting their DNA into the cells. This viral DNA hijacks the host cells machinery for reproduction of viral progeny, resulting in the host cells death. To overcome the viral infection, many bacteria and archaea have evolved several mechanisms. A major protective mechanism involves the use of restriction enzymes to degrade the invading viral DNA by cleaving it at specific restriction sites. At the same time, the host cell protects its own DNA from being cleaved by employing other enzymes called methylases, which methylate adenine or cytosine bases within host recognition sequences. For each of the restriction enzyme, the host cell produces a corresponding methylase that methylates and protects the ...
DNA Restriction Enzymes from Takara such as EcoO109I are high-quality: perform restriction enzyme digestion with reliable restriction endonucleases
This chapter summarizes the recent findings of bacterial genomics and comments on the themes and trends which are emerging. A variety of techniques and methods are available to construct physical maps, and those most commonly employed involve pulsed field gel electrophoresis (PFGE) of macrorestriction fragments generated by digesting intact genomic DNA, immobilized in agarose plugs, with rare-cutting enzymes. Hybridization techniques are often used to construct a map and to deduce the positions of genetic markers. In recent years significant effort has been devoted to developing direct-mapping techniques for large DNA molecules that do not require gel electrophoresis. Among the more promising of these are two new methods known as DNA combing and optical mapping, both of which make use of fluorescence microscopy and image analysis to visualize single DNA molecules. Overall, bacterial genomes range in size from about 0.6 to 9.4 Mb. In a recent review, it was suggested that there may be a relationship
A restriction endonuclease having one DNA binding site is proposed, synthesized from a restriction endonuclease that has one C-terminal domain and one N-terminal domain and two DNA binding sites, by proteolytic cleavage into the two domains or by cloning the gene segment that codes for the domains and expression of the domains and selection of the endonucleolytic domains having one DNA binding site. In addition, a method of synthesis of the restriction endonuclease and its use are claimed.
Today while Gracyn and Great Grandpa played outside, she dropped her pacifier near a squirrel and he ran off. While Gracyn watched the squirrel, Grandpa snatched up the paci. He told her that the squirrel took her paci. Gracyn believed him and took her nap today with no paci! Mommy scoured the house to find and hide all the pacifiers...even the one stuck to the end of her elephants trunk that she sleeps with every night. Tonight at bedtime, she asked for her favorite pillow and elephant.....when she noticed the pacifier was missing she looked up at Mommy and asked, "Squirrel took the paci?", when Mommy said yes, Gracyn said, "Look in my ear!" (G.Grandpa always "pulled" a paci out of her ear, like magic")!!!! Mommy reminded her that the squirrel took all of her pacifiers. This seemed to satisfy her. GRACYN WENT TO SLEEP ON HER OWN WITH NO PACIFER ...
Program: recoder # Rundate: Tue 15 Jul 2008 12:00:00 # Commandline: recoder # -sequence tembl:x65923 # -enzymes EcoRII # Report_format: table # Report_file: x65923.recoder ######################################## #======================================= # # Sequence: X65923 from: 1 to: 518 # HitCount: 34 # # KEY: # Dir: Direction (Rev for reverse complement) # EnzymeName: Enzyme name # RS-Pattern: Restriction enzyme recognition site pattern # Base-Posn: Position of base to be mutated # AAs: Amino acid. Original sequence(.)After mutation # Mutation: The base mutation to perform # # Creating silent mutations # #======================================= Start End Strand Dir EnzymeName RS-Pattern Base-Posn AAs Mutation 77 81 + . EcoRII CCWGG 78 P.P C-,G 77 81 + . EcoRII CCWGG 78 P.P C-,A 77 81 + . EcoRII CCWGG 78 P.P C-,T 77 81 + . EcoRII CCWGG 79 R.R A-,C 77 81 + . EcoRII CCWGG 81 R.R G-,A 107 111 + . EcoRII CCWGG 108 A.A C-,G 107 111 + . EcoRII CCWGG 108 A.A C-,A 107 111 + . EcoRII CCWGG 108 A.A ...
References[edit] ^ Roberts RJ (Nov 1976). "Restriction endonucleases". CRC Critical Reviews in Biochemistry. 4 (2): 123-64. doi:10.3109/10409237609105456. PMID 795607. ^ a b Kessler C, Manta V (Aug 1990). "Specificity of restriction endonucleases and DNA modification methyltransferases a review (Edition 3)". Gene. 92 (1-2): 1-248. doi:10.1016/0378-1119(90)90486-B. PMID 2172084. ^ Pingoud A, Alves J, Geiger R (1993). "Chapter 8: Restriction Enzymes". In Burrell M. Enzymes of Molecular Biology. Methods of Molecular Biology. 16. Totowa, NJ: Humana Press. pp. 107-200. ISBN 0-89603-234-5. ^ a b Arber W, Linn S (1969). "DNA modification and restriction". Annual Review of Biochemistry. 38: 467-500. doi:10.1146/annurev.bi.38.070169.002343. PMID 4897066. ^ Krüger DH, Bickle TA (Sep 1983). "Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts". Microbiological Reviews. 47 (3): 345-60. PMC 281580 . PMID 6314109. ^ Kobayashi I (Sep 2001). ...
These CRISPR/Cas9 knock-in mice constitutively express CRISPR associated protein 9 (|i|cas9|/i|) endonuclease and EGFP in a widespread fashion under the direction of a CAG promoter. When used in combination with single guide RNAs, they allow editing of single or multiple mouse genes |i|in vivo|/i| or |i|ex vivo|/i|.
NEB scientists continue to improve our portfolio of restriction enzymes, as well as explore their utility in new technologies.
Thanks, thats what I thought. Indeed, I thought youd reference the more interesting and relevant problem that you cant decide whether an array index is in the proper range. I rest my case. -- Matthias , From: Michael Norrish ,[email protected], , Content-Type: text/plain; charset=us-ascii , Date: Sat, 29 Dec 2001 10:51:01 +0000 , Cc: [email protected], [email protected], [email protected] , Reply-To: [email protected] , Sender: Michael Norrish ,[email protected], , , Matthias Felleisen writes: , , , Excuse my ignorance, but is the Standards notion of "conforming program" a , , decidable notion? If so, is your semantics accompanied by an algorithm? , , No. A strictly conforming program isnt allowed to divide by zero , (among many other restrictions) because this represents undefined , behaviour. , , Theres no way of deciding if a program is or isnt going to divide by , zero, so the class is undecidable. , , Michael ...
The graphic displays domains and Protease cut sites on the protein sequence. Drag your mouse right/left over the graphic. Use the selection boxes on the right to select which annotations to view simultaneously. Combine annotation with multiple checkmarks.. ...
The graphic displays domains and Protease cut sites on the protein sequence. Drag your mouse right/left over the graphic. Use the selection boxes on the right to select which annotations to view simultaneously. Combine annotation with multiple checkmarks.. ...
Sticky to Blunt end (Hypothetical) - posted in Genetics and Genomics: Hello, I have a hypothetical question. If I use a restriction enzyme, such as HinDIII on a plasmid vector to create sticky ends, is there a way to convert the overhangs to blunt ends without using Klenow fragment? There are restriction enzymes from NEB which recognize sequences of A and T only which would cleave the 3 overhang (like HpaI). Would this even work, or are my fundamentals lacking?
20 De Maio Na vossa paciência possuí as vossas almas, Luc.21.19 Depois que alguém nasce de novo e durante algum tempo seu pensamento e seu raciocínio perdem o vigor anterior. Temos que aprender a desenvolver a expressão da nova vida que é o formar da mente de Cristo em nós, Fil.2:5. Na vossa paciência possuí…
Linear) (Six-base) MAPSORT of: KIEE0945.seq Check: 8372 from: 1 to: 1050 KIEE0945.orf With 182 enzymes: SgfI * April 24, 2009 13:19 .. AcuI CTGAAGnnnnnnnnnnnnnn_nn Cuts at: 0 162 271 645 1050 Size: 162 109 374 405 Fragments arranged by size: 405 374 162 109 AflIII ACryG_T Cuts at: 0 396 518 1050 Size: 396 122 532 Fragments arranged by size: 532 396 122 AlfI GCAnnnnnnTGCnnnnnnnnnn_nn Cuts at: 0 329 1050 Size: 329 721 AloI GAACnnnnnnTCCnnnnnnn_nnnnn Cuts at: 0 694 726 1050 Size: 694 32 324 Fragments arranged by size: 694 324 32 AlwNI CAG_nnnCTG Cuts at: 0 375 645 1050 Size: 375 270 405 Fragments arranged by size: 405 375 270 AseI ATTA_AT Cuts at: 0 566 1050 Size: 566 484 AvaI CyCGr_G Cuts at: 0 41 1050 Size: 41 1009 BanI GGyrC_C Cuts at: 0 48 1050 Size: 48 1002 BbsI GAAGACnnnnnn_ Cuts at: 0 634 1050 Size: 634 416 BciVI GTATCCnnnnn_n Cuts at: 0 999 1050 Size: 999 51 BclI TGATC_A Cuts at: 0 74 1050 Size: 74 976 BglI GCCn_nnnnGGC Cuts at: 0 590 1050 Size: 590 460 Bme1580I G_kGCmC Cuts ...
Cumpara rapid si simplu Glucozamina Clorhidrat. Glucozamina Clorhidrat ajuta la reducerea durerilor articulare in Artrita, Reumatism, Artroza sau Coxartroza.
MmeI from Methylophilus methylotrophus belongs to the type II restriction-modification enzymes. It recognizes an asymmetric DNA sequence, 5-TCCRAC-3 (R indicates G or A), and cuts both strands at fixed positions downstream of the specific site. This particular feature has been exploited in transcript profiling of complex genomes (using serial analysis of gene expression technology). We have shown previously that the endonucleolytic activity of MmeI is strongly dependent on the presence of S-adenosyl-l-methionine (J. Nakonieczna, J. W. Zmijewski, B. Banecki, and A. J. Podhajska, Mol. Biotechnol. 37:127-135, 2007), which puts MmeI in subtype IIG. The same cofactor is used by MmeI as a methyl group donor for modification of an adenine in the upper strand of the recognition site to N(6)-methyladenine. Both enzymatic activities reside in a single polypeptide (919 amino acids [aa]), which puts MmeI also in subtype IIC of the restriction-modification systems. Based on a molecular model, generated ...
TY - JOUR. T1 - Inhibition of sequence-specific protein-DNA interaction and restriction endonuclease cleavage via triplex stabilization by poly(L-lysine)-graft-dextran copolymer. AU - Ferdous, Anwarul. AU - Akaike, Toshihiro. AU - Maruyama, Atsushi. PY - 2000/6. Y1 - 2000/6. N2 - Triplex stabilization by poly(L-lysine)-graft-dextran copolymer within a mammalian gene promoter inhibits the DNA binding activity of nuclear proteins from HeLa cells as well as restriction endonuclease cleavage at physiological pH and ionic conditions in vitro. Electrophoretic mobility shift assays using a 30-mer hornopurine-homopyrimidine stretch (located between -170 and -141 bp) of rat α1 (I) collagen gene promoter reveal that the copolymer, at its wide range of charge ratio with DNA, stabilizes triplex DNA and enhances triplex-specific inhibition of the protein - DNA interaction. When the triplex-forming region (located between -165 and -146 bp) of the promoter is engineered at the Bam H1 and Pst 1 sites of a ...
293472242 - EP 0985730 A1 2000-03-15 - Method for cloning and producing the snaBI restriction endonuclease and purification thereof - The methylase selection method was used to clone the SnaBI methylase gene (snaBIM) from Sphaerotilus natans (ATCC 15291). An active SnaBI methylase was cloned in E. coli using pSnaBI-2, a pUC19 derivative containing two SnaBI sites. Because methylase and restriction genes are usually located alongside each other in restriction-modification systems, efforts were made to clone the downstream DNA by inverse PCR. The missing portion of the SnaBI endonuclease was cloned by inverse PCR. A control, or C, protein was identified using the same technique. The two open reading frames snaBIR (ORF1) and snaBIC (ORF2) were orientated towards the SnaBI methylase gene (ORF). Attempts to establish a snaBIR-recombinant plasmid expressing the SnaBI endonuclease in E.coli modified with SnaBI methylase failed. Overexpression of the SnaBI endonuclease in E. coli required the use of the