Last data update: 31 October 2020 04:21 CET
Plasmid name: pCAGGShBcl-2 (LMBP 4633)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human B-cell leukemia/lymphoma 2α cDNA (BCL2, Bcl-2)
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Parental clone:||pCAGGS; pCRhBCL2|
|Further information:||The plasmid was constructed by inserting the human BCL2 cDNA as an EcoRI fragment from pCRhBCL2 into the EcoRI digested pCAGGS vector.
The orientation of the inserted gene, sense relative to the AG promoter, was confirmed by a BamHI digestion.
pCAGGShBcl-2 is useful for highly efficient expression of human BCL2 under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727135.1.
The human BCL2 EcoRI fragment was sequenced at the Department of Biomedical Molecular Biology (Ghent University, Belgium) and found identical to the DNA sequence described in the EMBL Nucleotide Sequence Database accession number BC027258.1, except for position 189 of the coding sequence, resulting in a silent mutation.
Other name of the plasmid is pCAGGSBcl-2.
|EMBL Accession number:||BC027258.1, view at EMBL, GenBank, DDBJ
LT727135.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||01/04/2003|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, HindII, NarI, PstI/PvuI and PvuII.|
|History of deposit:||This plasmid was deposited by Dr P. Vandenabeele(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Denecker et al., Cell Death Differ. 8 (2001), 829-840 [PMID: 11526436]
|Related plasmid reference:||Droga-Mazovec et al., J. Biol. Chem. 283 (2008), 19140-19150 [PMID: 18469004] [DOI: 10.1074/jbc.M802513200]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGShBcl-2 (LMBP 4633) is available at BCCM/GeneCorner. This plasmid was deposited by Dr P. Vandenabeele and was published in Denecker et al., 2001.|
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.