Last data update: 28 October 2020 04:17 CET
Plasmid name: pCAGGS-EhCdc42-DN (LMBP 4661)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human cell division cycle 42 GTP binding protein cDNA (CDC42, cdc42); dominant negative T17N 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 T17N mutant of the human CDC42 cDNA was amplified (PCR) and cloned, in frame with an N-terminal E-tag, into pCAGGS that was cut with NotI and XmaI.
pCAGGS-EhCdc42-DN is useful for highly efficient expression of the dominant negative mutant of human Cdc42 under the control of the AG promoter and the hCMV-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 LT727153.1.
The nucleotide sequence of the wt CDC42 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number BC003682.1. In accord with the depositor's data, codon 190 (CTG; Leu) was replaced by 'GTG' (Val).
Other name of the plasmid is pCAGGSE-hCdc42DN.
|EMBL Accession number:||BC003682.1, view at EMBL, GenBank, DDBJ
LT727153.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||09/10/2008|
Val Gly Lys Thr Cys Leu Leu wild type human CDC42 cDNA: 5'... gtt.ggt.aaa.aca.tgt.ctc.ctg ... 3' 17 Val Gly Lys Asn Cys Leu Leu mutated human CDC42 cDNA: 5'... gtt.ggt.aaa.aac.tgt.ctc.ctg ... 3' 17
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, Eco52I, NcoI, NotI and XhoI.|
|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-EhCdc42-DN (LMBP 4661) 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.