Last data update: 24 January 2024 16:39 CET
Plasmid name: pRSV-RBP3 (LMBP 5079)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p5079.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human recombination signal binding protein Jκ cDNA (RBP-Jκ); transcript variant hRBP3 |
| Promoter: | Rous sarcoma virus long terminal repeat (RSV-LTR) Phage SP6 promoter Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pRSV; pCMV/RBP3 |
| Further information: | The plasmid was constructed as follows: 1) the BglII-HindIII fragment of pcDNA3, containing the human cytomegalovirus immediate early promoter (CMV-IE) and enhancer, was replaced by the corresponding BglII-HindIII fragment of pRc/RSV, containing the RSV-LTR promoter, resulting in the pRSV vector. 2) the HindIII-XhoI fragment of pCMV/RBP3 (gift from Dr Israel, Institut Pasteur, Paris, France), containing the coding sequence of the transcript variant hRBP3 of the human recombination signal binding protein Jκ (hRBP-Jκ), was inserted into the HindIII-XhoI opened pRSV vector. The plasmid was designed for the expression of hRBP3 under the control of the RSV-LTR promoter in mammalian cells. There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin. The nucleotide sequences of pcDNA3 and pRc/RSV were obtained from Invitrogen (http://www.invitrogen.com; version B and version A respectively). The forward primer 5' cgtggatccGCATGGATTAAAAGGAAA 3' corresponding to the 5' end of the hRBP3 isoform ('aPCR3' in Amakawa et al., see Genbank (Accession number L07876.1)) and the reverse primer 5' cgcggatccTTAGGATACCACTGTTGGTGTAGA 3' were used in Brou et al. to isolate the full-length hRBP3 cDNA. The nucleotide sequence of aPCR3 has also been described in Genbank as the transcript variant 4 of hRBP-Jκ (Accession number NM_203284.1). Other name of the plasmid is pRSV-RBP wt. |
| EMBL Accession number: | L07876.1, view at EMBL, GenBank, DDBJ NM_203284.1, view at GenBank |
| Latest sequence update: | 13/10/2005 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI/NotI, BglII/HindIII, EcoRI/PstI, HindIII/XhoI, NcoI, NdeI and PvuII/XmnI. As compared to the compiled nucleotide sequence, the restriction site analysis pattern revealed that NcoI only cuts two times instead of three times. The NcoI site at position 665, at the start of the hRBP3 coding sequence, could not be experimentally confirmed. The compiled nucleotide sequence has not been adapted accordingly. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed in the research unit of Em. Prof. Dr G. Haegeman(2) by Dr W. Vanden Berghe(2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Plaisance et al., Mol. Cell. Biol. 17 (1997), 3733-3743 [PMID: 9199307] |
| Related plasmid reference: | Brou et al., Genes Dev. 8 (1994), 2491-2503 [PMID: 7958912] Amakawa et al., Genomics 17 (1993), 306-315 [PMID: 8406481] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pRSV-RBP3 (LMBP 5079) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Plaisance et al., 1997. |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.