The ssb gene, coding for single-stranded-DNA-binding protein (SSB), was cloned from four marine Shewanella strains that differed in their temperature and pressure optima and ranges of growth. All four Shewanella ssb genes complemented Escherichia coli ssb point and deletion mutants, with efficiencies that varied with temperature and ssb gene source. The Shewanella SSBs are the largest bacterial SSBs identified to date (24.9-26.3 kDa) and may be divided into conserved amino- and carboxy-terminal regions and a highly variable central region. Greater amino acid sequence homology was observed between the Shewanella SSBs as a group (72-87%) than with other bacterial SSBs (52-69%). Analysis of the amino acid composition of the Shewanella SSBs revealed several features that could correlate with pressure or temperature adaptation. SSBs from the three low-temperature-adapted Shewanella strains were an order of magnitude more hydrophilic than that from the mesophilic strain, and differences in the distribution of
Genome replication and maintenance occurs through the collective action of proteins that operate on single-stranded DNA (ssDNA). All cells express single-stranded DNA binding proteins (SSBs), which prevent errors by sequestering ssDNA with high-affinity, keeping it free from transient structures and protecting it from unwanted chemical modification. SSBs must be easily repositioned, or else risk stalling DNA replication and repair processes. How does a protein simulataneously bind DNA tightly yet diffuse rapidly?. Through a set of extensive all-atom molecular dynamics (MD) simulations, we have elucidated the molecular mechanism of SSB association with ssDNA. First, we showed that the same SSB-ssDNA complex can both spontaneously rearrange its structure and maintain its stable conformation depending on whether it is surrounded by physiological solution or a protein-crystal environment. Next, we probed the local interaction between ssDNA and SSB through simulations of mechanical unraveling of the ...
The present disclosure relates to methods for generating single-stranded DNA molecules of defined sequence and length. Specifically, a region of template containing target sequence is amplified by PCR or RCA, exogenous sequence is introduced by primers or probes used in amplification, double-stranded amplification products are converted to single-stranded amplification products, and single-stranded amplification products are trimmed to produce short single-stranded DNA molecules of defined sequence and length.
This article proves the existence of a hyper-precise global numerical meta-architecture unifying, structuring, binding and controlling the billion triplet codons constituting the sequence of single-stranded DNA of the entire human genome. Beyond the evolution and erratic mutations like transposons within the genome, its as if the memory of a fossil genome with multiple symmetries persists. This recalls the
Single-stranded DNA-binding proteins (SSBs), including replication protein A (RPA) in eukaryotes, play a central role in DNA replication, recombination, and repair. SSBs utilise an oligonucleotide/oligosaccharide-binding (OB) fold domain to bind DNA, and typically oligomerise in solution to bring multiple OB fold domains together in the functional SSB. SSBs from hyperthermophilic crenarchaea, such as Sulfolobus solfataricus, have an unusual structure with a single OB fold coupled to a flexible C-terminal tail. The OB fold resembles those in RPA, whilst the tail is reminiscent of bacterial SSBs and mediates interaction with other proteins. One paradigm in the field is that SSBs bind specifically to ssDNA and much less strongly to RNA, ensuring that their functions are restricted to DNA metabolism. Here, we use a combination of biochemical and biophysical approaches to demonstrate that the binding properties of S. solfataricus SSB are essentially identical for ssDNA and ssRNA. These features may ...
The present invention relates to a method of forming a three-stranded DNA molecule wherein each strand of the three-stranded DNA molecule is hybridized (that is, non-covalently bound) to at least one other strand of the three-stranded DNA molecule. The method comprises:contacting a recombination protein with a double-stranded DNA molecule and with a single-stranded DNA molecule sufficiently complementary to one strand of the double-stranded DNA molecule to hybridize therewith, which contacting is effected under conditions such that the single-stranded DNA molecule hybridizes to the double-stranded molecule so that the three stranded DNA molecule is formed.
The 10 bp DNA Ladder consists of thirty-three 10-bp repeats plus a fragment at 1668 bp and is suitable for sizing both double-stranded and single-stranded DNA fragments from 10 bp to 200 bp. The 100-bp band is approximately two to three times brighter than other ladder bands to provide internal orientation. In addition, because both DNA strands are of the same nucleotide composition, this product can be denatured to produce a set of single-stranded oligonucleotides increasing in length by 10-nucleotide increments. The double-stranded ladder can be visualized on 4% to 5% agarose gels after ethidium bromide staining. The single-stranded ladder can be visualized on an 8% urea-polyacrylamide gel after electrophoresis. This ladder can be easily radiolabeled using T4 polynucleotide kinase. ...
(CA2920328)This nucleic acid-encapsulating polymer micelle complex is characterized in being formed of: a block copolymer containing an uncharged hydrophilic polymer chain block and a cationic polymer chain block; and two single-stranded DNA molecules comprising mutually complementary base sequences of 1000 or more bases in length, double-stranded DNA of 1000 or more base pairs in length in which at least a part of the double helix structure has dissociated and taken on a single-stranded structure, or one single-stranded DNA molecule of 1000 or more bases in length.
(CN105451719)This nucleic acid-encapsulating polymer micelle complex is characterized in being formed of: a block copolymer containing an uncharged hydrophilic polymer chain block and a cationic polymer chain block; and two single-stranded DNA molecules comprising mutually complementary base sequences of 1000 or more bases in length, double-stranded DNA of 1000 or more base pairs in length in which at least a part of the double helix structure has dissociated and taken on a single-stranded structure, or one single-stranded DNA molecule of 1000 or more bases in length.
Single‐stranded DNA (ssDNA) regions form as an intermediate in many DNA‐associated transactions. Multiple cellular proteins interact with ssDNA via the oligonucleotide/oligosaccharide‐binding (OB) fold domain. The heterotrimeric, multi‐OB fold domain‐containing Replication Protein A (RPA) complex has an essential genome maintenance role, protecting ssDNA regions from nucleolytic degradation and providing a recruitment platform for proteins involved in responses to replication stress and DNA damage. Here, we identify the uncharacterized protein RADX (CXorf57) as an ssDNA‐binding factor in human cells. RADX binds ssDNA via an N‐terminal OB fold cluster, which mediates its recruitment to sites of replication stress. Deregulation of RADX expression and ssDNA binding leads to enhanced replication fork stalling and degradation, and we provide evidence that a balanced interplay between RADX and RPA ssDNA‐binding activities is critical for avoiding these defects. Our findings establish ...
This article proves the existence of a hyper-precise global numerical meta-architecture unifying, structuring, binding and controlling the billion triplet codons comprising the sequence of single-stranded DNA of the entire human genome. Beyond the evolution and erratic mutations like transposons within the genome, its as if the memory of a fossil genome with multiple symmetries persists. This recalls the "intermingling" of information characterizing the fractal universe of chaos theory. The result leads to a balanced and perfect tuning between the masses of the two strands of the huge DNA molecule that constitute our genome. We show here how codon populations forming the single-stranded DNA sequences can constitute a critical approach to the understanding of junk DNA function. Then, we suggest revisiting certain methods published in our 2009 book "Codex Biogenesis". In fact, we demonstrate here how the universal genetic code table is a powerful analytical filter to characterize single-stranded ...
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In ,9209101632.AA03158 at anolis.bnr.usu.edu, thsiao at ANOLIS.BNR.USU.EDU writes: , Dear Netters, , , I am interested in the use of PCR enhancers. I would like to know if any one , has experience with the use of Stratagenes Perfect Match Polymerase Enhancer. , Is this a single-stranded DNA binding protein? What is the difference with , USBs single-stranded DNA binding protein or GP 32? How effective are these , products in getting better PCR yields? Any suggestion and tips will be , appreciated. , , +---------------Ting H. Hsiao ---------------+ , , Dept. of Biology, Utah State University , , , Logan, UT 84322-5303, U.S.A. , , , PHONE: (801) 750-2549; FAX: (801) 750-1575 , , , E-MAIL: thsiao at anolis.bnr.usu.edu , , +--------------------------------------------+ An issue of Biotechniques (I think, not absolutely sure) a couple of years ago revealed that Stratagenes Perfect Match is actually a 5x10E-5 M solution of tetra-methyl-ammonium chloride. ...
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ssDNA Antibody is an affinity-purified IgG that recognizes ssDNA. The ssDNA Antibody was raised in rabbit and can be used for IHC detection of apoptotic cells.
The effect of various DNA dyes and enhancers on ssDNA fluorescence and dsDNA melting temperature. (A) TNF-1 oligonucleotide (ssDNA, 45.5% GC; 1 μM final concen
Imetajatest on mitokondriaalsed nukleoidid kõige paremini kirjeldatud inimeses. Samuti on nukleoidid leitud roti, rohepärdiku ja potoroo maksarakkude mitokondritest[6][7]. Imetajate mtDNA paiknemine diskreetsete üksustena oli esmakordselt näidatud 1991. aastal inimrakkude mtDNA värvimisel DAPI-ga[8] ning see oli hiljem kinnitatud ka immunofluorestsentsi katsetes, kus kasutati anti-DNA antikehi[9][10]. mtDNA kompleksid valkudega olid imetajates kirjeldatud siis, kui näidati mtDNA seondumist DNA helikaasiga Twinkle inimrakkudes[11]. Seejärel demonstreeriti mitme uuringu käigus ka teiste valkude esinemine mtDNA-ga, peaaegu alati esinesid koos mtDNA-ga mitokondriaalne transkriptsioonifaktor A (TFAM, ingl. mitochondrial transcription factor A), pagaripärmi Apf2p valgu homoloog, ning mitokondriaalne üheahelalise DNA-ga seonduv valk mtSSB (ingl. mitochondrial single strand binding protein), mis on pagaripärmi Rim1p homoloog. TFAM on seejuures valk, mille põhiline funktsioon seisneb mtDNA ...
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
1AE3: Analyses of the stability and function of three surface mutants (R82C, K69H, and L32R) of the gene V protein from Ff phage by X-ray crystallography.
1GVP: Analyses of the stability and function of three surface mutants (R82C, K69H, and L32R) of the gene V protein from Ff phage by X-ray crystallography.
... definition, a single-stranded molecule of RNA that is synthesized in the nucleus from a DNA template and then enters the cytoplasm, where its genetic code specifies the amino acid sequence for protein synthesis. See more.
Gene target information for Ssbp3 - single-stranded DNA binding protein 3 (house mouse). Find diseases associated with this biological target and compounds tested against it in bioassay experiments.
WANGNN at UCBEH.SAN.UC.EDU wrote: : Hi Netters: : I am going run the gel retardation to show the band shifting using my protein : and a small fragment DNA. The small fragment is about 20 bases, which is : planned to be synthesized. For sure, the synthesis of a 20-mer of : oligonucleotide is much cheaper than that of two strands to make a ds DNA. : My question is if some methods can be used to synthesis such small ds DNA : fragment (20bp) from the single strand oligonucleotide. : Any suggestion will be appreciated. Dear Hong, All DNA polymerases require a short primer to initiate synthesis. Even RNA polymerases require a promoter site. If you were really clever, you could design a DNA hairpin structure which would fold back on itself to give you a double-stranded region (really just a big stem-loop). You could use the Zucker folding program (available on several computer platforms) to help predict the folding, and you would have to be sure to anneal at low concentration to avoid intermolecular ...
Reverse transcriptases can synthesize a complementary DNA strand initiating from a primer using RNA (cDNA synthesis) or single-stranded DNA as a template.
S1 nuclease (EC 3.1.30.1, ~300 aa) is a fungal nuclease that degrades single-stranded nucleic acids and is preferentially active against DNA. Used experimentally to analyse the structure of DNA:RNA hybrids (S1 nuclease mapping, Berk-Sharp technique), and to remove single-stranded extensions from DNA to produce blunt ends (see restriction endonucleases). ...
Integrated DNA Technologies (IDT) is a company that manufactures and sells oligonucleotides (short, single-stranded DNA or RNA molecules). On their website, they provide the Oligo Analyzer service, which can be used to examine an oligonucleotide (oligo) in a variety of ways. This service is primarily available through a web interface, but the Analyze, Hetero-Dimer, and Self-Dimer functions are also available as SOAP functions. The Analyze service returns physical properties of a given oligo sequence. The Hetero-Dimer examines possible duplexes when one oligo is combined with another. The Self-Dimer reports possible duplexes and their stabilities when an oligo hybridizes with itself ...
For example, to join together two BioBricks, you would first cut both plasmids with restriction enzymes, turning one into an "insert" by getting rid of the rest of the plasmid, and turning the other into a "vector" by opening a space in the plasmid in front of the BioBrick. Because As always pair with Ts and Gs always pair with Cs, the overhanging edges of single-stranded DNA that your restriction enzymes left behind will match up to make double stranded DNA. You then mix together the insert and vector with a special enzyme called a "ligase" that can join together two broken pieces of DNA. The result is a composite plasmid that contains two BioBricks, now side by side[4][5] [6]. It is important to note that this new larger composite part has the same restriction sites as the smaller parts it was originally made from. This is what is meant by preserving "key structural elements" that allow one component of any size to be easily connected to any other component[1]. Also note that the "scar" ...
The first model describes the dynamic behavior of the virus production in terms of the law of mass action (Sidorenko & Reichl 2004). Therefore the temporal changes in concentrations of titrated plasmids, synthesized proteins, replicated single-stranded DNA and formed virus capsids and finally concentration of viral particles are simulated using MathWorks® MATLAB R2010b ...
CHEFSCHOICE Trizor XV EdgeSelect Sharpener: 3-stage process; combines the power of the triple-bevel Trizor edge with 15-degree, EdgeSelect technology; converts 20-degree edges, as well as double- and single-bevel edges, into Trizor edges
Traditionally, recombination reactions promoted by RecA-like proteins initiate by forming a nucleoprotein filament on a single-stranded DNA (ssDNA), which then pairs with homologous double-stranded DNA (dsDNA). In this paper, we describe a novel pairing process that occurs in an unconventional manner: RecA protein polymerizes along dsDNA to form an active nucleoprotein filament that can pair and exchange strands with homologous ssDNA. Our results demonstrate that this inverse reaction is a unique, highly efficient DNA strand exchange reaction that is not due to redistribution of RecA protein from dsDNA to the homologous ssDNA partner. Finally, we demonstrate that the RecA protein-dsDNA filament can also pair and promote strand exchange with ssRNA. This inverse RNA strand exchange reaction is likely responsible for R-loop formation that is required for recombination-dependent DNA replication.
Using various replication mutants of E. coli, the host genes that participate in the replication of some K12-specific single-stranded DNA phages have been
Eukaryotic cells encode two homologs of Escherichia coli RecA protein, Rad51 and Dmc1, which are required for meiotic recombination. Rad51, like E.coli RecA, forms helical nucleoprotein filaments that promote joint molecule and heteroduplex DNA formation. Electron microscopy reveals that the human meiosis-specific recombinase Dmc1 forms ring structures that bind single-stranded (ss) and double-stranded (ds) DNA. The protein binds preferentially to ssDNA tails and gaps in duplex DNA. hDmc1-ssDNA complexes exhibit an irregular, often compacted structure, and promote strand-transfer reactions with homologous duplex DNA. hDmc1 binds duplex DNA with reduced affinity to form nucleoprotein complexes. In contrast to helical RecA/Rad51 filaments, however, Dmc1 filaments are composed of a linear array of stacked protein rings. Consistent with the requirement for two recombinases in meiotic recombination, hDmc1 interacts directly with hRad51.. ...
Mutations in genes encoding components of the mitochondrial DNA (mtDNA) replication machinery cause mtDNA depletion syndromes (MDSs), which associate ocular features with severe neurological syndromes. Here, we identified heterozygous missense mutations in single-strand binding protein 1 (SSBP1) in 5 unrelated families, leading to the R38Q and R107Q amino acid changes in the mitochondrial single-stranded DNA-binding protein, a crucial protein involved in mtDNA replication. All affected individuals presented optic atrophy, associated with foveopathy in half of the cases. To uncover the structural features underlying SSBP1 mutations, we determined a revised SSBP1 crystal structure. Structural analysis suggested that both mutations affect dimer interactions and presumably distort the DNA-binding region. Using patient fibroblasts, we validated that the R38Q variant destabilizes SSBP1 dimer/tetramer formation, affects mtDNA replication, and induces mtDNA depletion. Our study showing that mutations in ...
In one method, DNA fragments, of approximately 2-200 bases in length, or deoxynucleotides (single bases), are administered topically to the epidermis, either in a liposome preparation or in another appropriate vehicle, such as propylene glycol, in a quantity sufficient to enhance melanin production. As used herein, "DNA fragments" refers to single-stranded DNA fragments, double-stranded DNA fragments, a mixture of both single-and double-stranded DNA fragments, or deoxynucleotides. "Deoxynucleotides" refers to either a single type of deoxynucleotide or a mixture of different deoxynucleotides. The DNA fragments or deoxynucleotides can come from any appropriate source. For example, salmon sperm DNA can be dissolved in water, and then the mixture can be autoclaved to fragment the DNA. The fragments can additionally be UV-irradiated. The liposome preparation can be comprised of any liposomes which penetrate the stratum corneum and fuse with the cell membrane, resulting in delivery of the contents of ...
Canonical single-stranded DNA-binding proteins (SSBs) from the oligosaccharide/oligonucleotide-binding (OB) domain family are present in all known organisms and are critical for DNA replication, recombination and repair. The SSB from the hyperthermophilic crenarchaeote Sulfolobus solfataricus (SsoSSB) has a simple domain organization consisting of a single DNA-binding OB fold coupled to a flexible C-terminal tail, in contrast with other SSBs in this family that incorporate up to four OB domains. Despite the large differences in the domain organization within the SSB family, the structure of the OB domain is remarkably similar all cellular life forms. However, there are significant differences in the molecular mechanism of ssDNA binding. We have determined the structure of the SsoSSB OB domain bound to ssDNA by NMR spectroscopy. We reveal that ssDNA recognition is modulated by base-stacking of three key aromatic residues, in contrast with the OB domains of human RPA and the recently discovered ...
Transfection of cells with gene-specific, single-stranded oligonucleotides can induce the targeted exchange of one or two nucleotides in the targeted gene. To characterize the features of the DNA-repair mechanisms involved, we examined the maximal distance for the simultaneous exchange of two nucleotides by a single-stranded oligonucleotide. The chosen experimental system was the correction of a hprt- point mutation in a hamster cell line, the generation of an additional nucleotide exchange at a variable distance from the first exchange position and the investigation of the rate of simultaneous nucleotide exchanges. The smaller the distance between the two exchange positions, the higher was the probability of a simultaneous exchange. The detected simultaneous nucleotide exchanges were found to cluster in a region of about fourteen nucleotides upstream and downstream from the first exchange position. We suggest that the mechanism involved in the repair of the targeted DNA strand utilizes only a short
TY - JOUR. T1 - Enzymatic production of single-stranded DNA as a target for fluorescence in situ hybridization. AU - van Dekken, H.. AU - Pinkel, D.. AU - Mullikin, J.. AU - Gray, J. W.. PY - 1988/9/1. Y1 - 1988/9/1. N2 - This study demonstrates that Exonuclease III (Exo III) can be used to produce sufficient single-stranded (ss)DNA in chromosomes and cells to allow in situ hybridization. In this study, all of the probes were modified with biotin and the probe binding was visualized with fluorescein-labeled avidin. Exo III digestion starting at naturally occurring breaks in methanol-acetic acid preparations produced enough ssDNA for strong hybridization when human genomic DNA was used to probe human chromosomes. Pretreatment with the endonucleases EcoRI, Hind III and BamHI was used to produce more sites for initiation of Exo III digestion when using a chromosome-specific repetitive probe specific to a small chromosomal subregion near the telomere of human chromosome 1(1p36). The fluorescence ...
The CRISPR/Cas9 system used in conjunction with single stranded DNA donors is revolutionising our ability to generate targeted mutations directly in the embryo. Whilst short synthetic DNA molecules facilitate this, the use of longer single-stranded DNA donors is a more recent addition to the genome editing toolbox. The two new articles summarised here compare long and short single-stranded donors in a high-throughput setting, both look at conditional knock-out mutants while also presenting advances for the generations of point mutations.. In the first study, led by researchers in Lydia Tebouls group at the MRC Harwell institute, long single-stranded molecules are utilised to facilitate the generation of conditional alleles. They also apply the system to the introduction of point mutations remote from the recognition site of active Cas9/sgRNA complexes, which up to now has not been possible. This last technique is particularly valuable for human genomic sequencing since it enhances our ability ...
In molecular biology, hybridization (or hybridisation) is a phenomenon in which single-stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) molecules anneal to complementary DNA or RNA. Though a double-stranded DNA sequence is generally stable under physiological conditions, changing these conditions in the laboratory (generally by raising the surrounding temperature) will cause the molecules to separate into single strands. These strands are complementary to each other but may also be complementary to other sequences present in their surroundings. Lowering the surrounding temperature allows the single-stranded molecules to anneal or "hybridize" to each other. DNA replication and transcription of DNA into RNA both rely upon nucleotide hybridization, as do molecular biology techniques including Southern blots and Northern blots, the polymerase chain reaction (PCR), and most approaches to DNA sequencing. Hybridization is a basic property of nucleotide sequences and is taken advantage of ...
The initiation of an immune response is dependent on the activation and maturation of dendritic cells after sensing pathogen associated molecular patterns by pattern recognition receptors. However, the response needs to be balanced as excessive pro-inflammatory cytokine production in response to viral or stress-induced pattern recognition receptor signaling has been associated with severe influenza A virus (IAV) infection. Here, we use an inhibitor of Toll-like receptor (TLR)3, a single-stranded oligonucleotide (ssON) with the capacity to inhibit certain endocytic routes, or a TLR3 agonist (synthetic double-stranded RNA PolyI:C), to evaluate modulation of innate responses during H1N1 IAV infection. Since IAV utilizes cellular endocytic machinery for viral entry, we also assessed ssONs capacity to affect IAV infection. We first show that IAV infected human monocyte-derived dendritic cells (MoDC) were unable to up-regulate the co-stimulatory molecules CD80 and CD86 required for T cell activation.
APOBEC3G is the best known of several DNA cytosine deaminases that function to inhibit the replication of parasitic genetic elements including the lentivirus HIV. Several high-resolution structures of the APOBEC3G catalytic domain have been generated, but none reveal how this enzyme binds to substrate single-stranded DNA. Here, we constructed a panel of APOBEC3G amino acid substitution mutants and performed a series of biochemical, genetic, and structural assays to distinguish between
Can catalyze the hydrolysis of ATP in the presence of single-stranded DNA, the ATP-dependent uptake of single-stranded DNA by duplex DNA, and the ATP-dependent hybridization of homologous single-stranded DNAs. It interacts with LexA causing its activation and leading to its autocatalytic cleavage.
Can catalyze the hydrolysis of ATP in the presence of single-stranded DNA, the ATP-dependent uptake of single-stranded DNA by duplex DNA, and the ATP-dependent hybridization of homologous single-stranded DNAs. It interacts with LexA causing its activation and leading to its autocatalytic cleavage.
Omics is a science that comprehensively embraces the four disciplines of genomics, transcriptomics, proteomics and metabolomics (Fig. 2), and has developed rapidly in the past decade triggered by improvements in genome decoding techniques and processing speed. These improvements have led to some remarkable milestones in genomic research, including sequencing of the complete genomes of the flowering plant Arabidopsis thaliana, rice and soy-bean.. Similarly, the DNA microarray technique used in transcriptome analysis, the analysis of RNA transcribed from DNA, has also improved dramatically over the past decade. In this technique, hundreds of thousands of single-stranded DNA fragments are fixed in holes or spots on a glass substrate, and fluorescently labeled RNA are dropped onto the substrate surface. RNA complementary to a DNA fragment will become bound to the DNA, which causes the combined compound to emit fluorescent light. From the fluorescence intensity of each spot, researchers can ...
Background Single-stranded DNA-binding (SSB) proteins play an essential role in all in vivo processes involving ssDNA. They interact with ssDNA and RNA, in an independent from sequence manner, preventing single-stranded nucleic acids from hybridization and degradation. by nucleases [1]. SSB proteins play a central role in DNA replication, repair and recombination [2-4]. They have been identified in all classes of organisms, performing similar functions but displaying little sequence similarity and very different ssDNA binding properties. Based on their oligomeric state, SSBs can be classified into four groups: monomeric, homodimeric, heterotrimeric and homotetrameric. A prominent feature of all SSBs is that the DNA-binding domain is made up of a conserved motif, the OB (oligonucleotide binding) ICG-001 chemical structure fold [5]. Most of the bacterial SSBs exist as homotetramers. However, recent discoveries have shown that. SSB proteins from the genera Thermus and Deinococcus possess a ...
For about 10 years until 2000, my labs research activities were focused on the mechanism of recombinational repair of double-strand breaks in DNA. We focused our efforts on two model systems: one involved the repair of restriction enzyme cleavages at specific mammalian chromosomal loci and the second explored the biochemical properties of purified yeast Rad51 protein, an essential catalyst for synapsing the broken ends of DNA with an intact homologue of that sequence. We also explored the roles of Rad52 and PRA (single-strand DNA binding protein) in the repair process.In 2000, I became Emeritus Professor in Biochemistry and stepped down from the Directorship of the Beckman Center. Much of my activities since then have been involved in writing a biography of the genetics pioneer George Beadle, published in 2003, plus articles for other publications elaborating on Beadles legacy for todays science. Over the years I have been and continue to be an activist in public policy issues affecting ...
Hypertrophic pancreatic islets (PI) of Goto Kakizaki (GK) diabetic rats contain a lower number of β-cells vs. non-diabetic Wistar rat PI. Remaining β-cells contain reduced mitochondrial (mt) DNA per nucleus (copy number), probably due to declining mtDNA replication machinery, decreased mt biogenesis or enhanced mitophagy. We confirmed mtDNA copy number decrease down to ,30% in PI of one-year-old GK rats. Studying relations to mt nucleoids sizes, we employed 3D superresolution fluorescent photoactivable localization microscopy (FPALM) with lentivirally transduced Eos conjugate of mt single-stranded-DNA-binding protein (mtSSB) or transcription factor TFAM; or by 3D immunocytochemistry ...
Double diffusion experiments with antibodies to single-stranded (s-s) DNA confirmed physicochemical data that adeno-associated satellite virions contain s-s DNA but the DNA extracted from such virions is double-stranded. Positive immunologic reactions of intact virions with sera from man and mouse with "autoimmune" disease and specifically immunized rabbits are consistent with the hypothetical role if viruses as immunogens or as the antigenic participants of immunecomplex vasculitis.. ...
Single-stranded DNA-binding protein required for homologous recombination in meiosis I. Required for double strand breaks (DSBs) repair and crossover formation and promotion of faithful and complete synapsis. Not required for the initial loading of recombinases but required to maintain a proper number of RAD51 and DMC1 foci after the zygotene stage. May act by ensuring the stabilization of recombinases, which is required for successful homology search and meiotic recombination. Displays Single-stranded DNA 3-5 exonuclease activity in vitro (By similarity ...