Last data update: 30 July 2021 10:22 CEST
Plasmid name: pCAGGS-EhRhoA-DP (LMBP 4658)
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|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
Human ras homolog family member A cDNA (RHOA, ARH12, ARHA, Rho12, RhoA, RHOH12); constitutive active Q63L 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 constitutive active Q63L 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-DP is useful for highly efficient expression of the constitutive active or dominant positive 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 LT727150.1.
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-hRhoADP.
|EMBL Accession number:||AF498970.1, view at EMBL, GenBank, DDBJ
LT727150.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||24/10/2006|
Thr Ala Gly Gln Glu Asp Tyr wild type hRHOA cDNA: 5'... aca.gct.ggg.cag.gaa.gat.tat ...3' 63 Thr Ala Gly Leu Glu Asp Tyr mutated hRHOA cDNA: 5'... aca.gct.ggg.ctc.gaa.gat.tat ...3' 63
|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-DP (LMBP 4658) 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.