DNA fragments on a single-molecule degree reveals patterns
BCAS2, a protein enriched in superior prostate most cancers, interacts with NBS1 to bolster DNA double-strand break restore
Background: Breast most cancers amplified sequence 2 (BCAS2) performs important roles in pre-mRNA splicing and androgen receptor transcription. Earlier analysis urged that BCAS2 is worried in double-strand breaks (DSB); on account of this truth, we aimed to characterise its mechanism and place in prostate most cancers (PCa).
Methods: Western blotting and immunofluorescence microscopy have been used to assay the roles of BCAS2 inside the DSBs of PCa cells and apoptosis in Drosophila, respectively. The impression of BCAS2 dosage on non-homologous end changing into a member of (NHEJ) and homologous recombination (HR) have been assayed by actual end-joining assay and transfer cytometry, respectively. Glutathione-S-transferase pulldown and co-immunoprecipitation assays have been used to seek out out whether or not or not and the way in which BCAS2 interacts with NBS1. The expression of BCAS2 and totally different proteins in human PCa was determined by immunohistochemistry.
Outcomes: BCAS2 helped restore radiation-induced DSBs successfully in every human PCa cells and Drosophila. BCAS2 enhanced every NHEJ and HR, most likely by interacting with NBS1, which involved the BCAS2 N-terminus as successfully as every the NBS1 N- and C-termini. The overexpression of BCAS2 was significantly associated to bigger Gleason and pathology grades and shorter survival in victims with PCa.
Conclusion: BCAS2 promotes two DSB restore pathways by interacting with NBS1, and it might impact PCa improvement.
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human CSP . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody for detection of CSP from Human, Mouse, Rat. This CSP antibody is for WB, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the C-terminal region of human CSP at AA range: 100-180
Description: A polyclonal antibody for detection of CSP from Human, Mouse, Rat. This CSP antibody is for WB, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the C-terminal region of human CSP at AA range: 100-180
Description: A polyclonal antibody for detection of CSP from Human, Mouse, Rat. This CSP antibody is for WB, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the C-terminal region of human CSP at AA range: 100-180
Description: DNAJC5 is a member of the J protein family. J proteins function in many cellular processes by regulating the ATPase activity of 70 kDa heat shock proteins. The encoded protein (DnaJ heat shock protein family (Hsp40) member C5) plays a role in membrane trafficking and protein folding, and has been shown to have anti-neurodegenerative properties. The encoded protein is known to play a role in cystic fibrosis and Huntington's disease. A pseudogene of DNAJC5 is located on the short arm of chromosome 8.
Description: DNAJC5 is a member of the J protein family. J proteins function in many cellular processes by regulating the ATPase activity of 70 kDa heat shock proteins. The encoded protein (DnaJ heat shock protein family (Hsp40) member C5) plays a role in membrane trafficking and protein folding, and has been shown to have anti-neurodegenerative properties. The encoded protein is known to play a role in cystic fibrosis and Huntington's disease. A pseudogene of DNAJC5 is located on the short arm of chromosome 8.
Description: DNAJC5 is a member of the J protein family. J proteins function in many cellular processes by regulating the ATPase activity of 70 kDa heat shock proteins. The encoded protein (DnaJ heat shock protein family (Hsp40) member C5) plays a role in membrane trafficking and protein folding, and has been shown to have anti-neurodegenerative properties. The encoded protein is known to play a role in cystic fibrosis and Huntington's disease. A pseudogene of DNAJC5 is located on the short arm of chromosome 8.
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human DNAJC5 / CSP (aa177-191). This antibody is tested and proven to work in the following applications:
Sequencing and Characterisation of Full Mitochondrial DNA Genome for Trigonopoma pauciperforatum (Cypriniformes: Cyprinidae: Danioninae) with Phylogenetic Consideration
The Trigonopoma pauciperforatum or the redstripe rasbora is a cyprinid usually current in marshes and swampy areas with slight acidic tannin-stained water inside the tropics. On this analysis, the entire mitogenome sequence of pauciperforatum was first amplified in two elements using two pairs of overlapping primers after which sequenced.
The size of the mitogenome is 16,707 bp, encompassing 22 swap RNA genes, 13 protein-coding genes, two ribosomal RNA genes and a putative administration space. An similar gene organisation was detected between this species and totally different family members.
The heavy strand accommodates 28 genes whereas the sunshine strand houses the remaining 9 genes. Most protein-coding genes utilise ATG as start codon except for COI gene which makes use of GTG as a substitute. The terminal associated sequence (TAS), central conserved sequence block (CSB-F, CSB-D and CSB-E) along with variable sequence block (CSB-1, CSB-2 and CSB-3) are conserved inside the administration space.
The utmost likelihood phylogenetic tree revealed the divergence of pauciperforatum from the basal space of the foremost clade, the place its evolutionary relationships with Boraras maculatus, Rasbora cephalotaenia and R. daniconius are poorly resolved as urged by the low bootstrap values.
This work contributes within the route of the genetic helpful useful resource enrichment for peat swamp conservation and full in-depth comparisons all through totally different phylogenetic researches accomplished on the Rasbora-related genus.
Reconstructing double-stranded DNA fragments on a single-molecule diploma reveals patterns of degradation in historic samples
Intensive manipulations involved inside the preparation of DNA samples for sequencing have hitherto made it unimaginable to seek out out the precise building of double-stranded DNA fragments being sequenced, such as a result of the presence of blunt ends, single-stranded overhangs, or single-strand breaks.
We proper right here describe MatchSeq, a method that mixes single-stranded DNA library preparation from diluted DNA samples with computational sequence matching, allowing the reconstruction of double-stranded DNA fragments on a single-molecule diploma.
The equipment of MatchSeq to Neanderthal DNA, a really superior provide of degraded DNA, reveals that 1- or 2-nt overhangs and blunt ends dominate the ends of historic DNA molecules and that temporary gaps exist, which can be predominantly introduced on by the dearth of specific individual purines.
We extra current that deamination of cytosine to uracil occurs in every single- and double-stranded contexts close to the ends of molecules, and that single-stranded elements of DNA fragments are enriched in pyrimidines. MatchSeq provides unprecedented choice for interrogating the buildings of fragmented double-stranded DNA and could possibly be utilized to fragmented double-stranded DNA isolated from any natural provide. The tactic relies on well-established laboratory strategies and would possibly merely be built-in into routine information period.
This danger is confirmed by the worthwhile reconstruction of double-stranded DNA fragments from beforehand revealed single-stranded sequence information, allowing a further full characterization of the biochemical properties not solely of historic DNA however as well as of cell-free DNA from human blood plasma, a clinically associated marker for the evaluation and monitoring of sickness.
Description: A DNA sequence encoding the mature variant of ov-VEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight Cysteine residues of the central Cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999; Wise et al., 1999). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E cannot bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Description: A DNA sequence encoding the mature variant of ovVEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999; Wise et al., 1999). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E can not bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Description: A DNA sequence encoding the mature variant of ovVEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999; Wise et al., 1999). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E can not bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Description: A DNA sequence encoding the mature variant of ov-VEGF-E isolate D1701 was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight Cysteine residues of the central Cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins. Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E cannot bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Recombinant Virus VEGF E (Orf Virus) Protein, His, E.coli-1mg
Description: A DNA sequence encoding the first 116 amino acid residue of Orf virus VEGF-E isolate D1701 (Dehio et al., 1999 EMBO J. 18:363-374; GenBank accession No. AF106020) was fused with a DNA sequence encoding to the C-terminal heparin binding domain of human VEGF165. The chimeric protein was expressed in insect cells using a baculovirus expression system. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994 J. Virol 68:84-92). Different isolates of orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appear to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999 EMBO J. 18:363-374; Wise et al., 1999 Proc. Natl. Acad. Sci USA 96:3071-3076). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E and hb-VEGF-E can not bind to VEGF receptor-1 (Flt-1). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor–2/ KDR. Compared to human VEGF165 this virus form has no heparin-binding domain and seems to be a freely secreted protein comparable to VEGF121. In order to compare this form with human VEGF165, an additional heparin-binding domain was engineered at the C-terminus to allow interaction with proteo-aminoglycans and heparan sulfate. These form is also able to interact with neuropillin–1.
Orf virus VEGF-E, Heparin-binding Recombinant Protein
Description: A DNA sequence encoding the first 116 amino acid residue of Orf virus VEGF-E isolate D1701 (Dehio et al., 1999 EMBO J. 18:363-374; GenBank accession No. AF106020) was fused with a DNA sequence encoding to the C-terminal heparin binding domain of human VEGF165. The chimeric protein was expressed in insect cells using a baculovirus expression system. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994 J. Virol 68:84-92). Different isolates of orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appear to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999 EMBO J. 18:363-374; Wise et al., 1999 Proc. Natl. Acad. Sci USA 96:3071-3076). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E and hb-VEGF-E can not bind to VEGF receptor-1 (Flt-1). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor–2/ KDR. Compared to human VEGF165 this virus form has no heparin-binding domain and seems to be a freely secreted protein comparable to VEGF121. In order to compare this form with human VEGF165, an additional heparin-binding domain was engineered at the C-terminus to allow interaction with proteo-aminoglycans and heparan sulfate. These form is also able to interact with neuropillin–1.
Description: A DNA sequence encoding the mature variant of ovVEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa.
