Last data update: 23 October 2020 04:27 CEST
Plasmid name: pCAGGS-EhRhoA-DN (LMBP 4657)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human ras homolog family member A cDNA (RHOA, ARH12, ARHA, Rho12, RhoA, RHOH12); dominant negative T19N mutant
|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
|Further information:||The plasmid was constructed as follows: the dominant negative T19N mutant of the human RHOA cDNA was amplified (PCR) and cloned, in frame with an N-terminal E-tag, into pCAGGS that was cut with NotI and XmaI.
pCAGGS-EhRhoA-DN is useful for highly efficient expression of the dominant negative mutant of human RhoA 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 LT727149.1.
The nucleotide sequence of the wt RHOA cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF498970.1.
Other name of the plasmid is pCAGGSE-hRhoADN.
|EMBL Accession number:||AF498970.1, view at EMBL, GenBank, DDBJ
LT727149.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||24/10/2006|
Cys Gly Lys Thr Cys Leu Leu wild type hRhoA cDNA: 5'... tgt.gga.aag.aca.tgc.ttg.ctc ...3' 19 Cys Gly Lys Asn Cys Leu Leu mutated hRhoA cDNA: 5'... tgt.gga.aag.aac.tgc.ttg.ctc ...3' 19
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, Eco52I and NotI.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by Dr P. Schotte(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Schotte et al., J. Biol. Chem. 279 (2004), 25134-25142 [PMID: 15060067]
|Related plasmid reference:||Sander et al., J. Cell Biol. 147 (1999), 1009-1022 [PMID: 10579721]
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-EhRhoA-DN (LMBP 4657) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Schotte et al., 2004.|
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.