Last data update: 30 October 2020 04:11 CET
Plasmid name: pCAGGS-E-hPELI1-PM (LMBP 5192)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human pellino 1 cDNA (PELI1); mutated sequence containing a double point mutation (H369S/C371S) in the RING-domain (PELI1-PM)
|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-E-hPELI1; pCAGGS|
|Further information:||The plasmid was constructed as follows: 1) a double point mutation H369S/C371S in the RING-domain of the hPELI1 cDNA was generated by PCR on pCAGGS-E-hPELI1; 2) the NotI/XhoI digested PCR amplicon was fused in frame to the E-tag of a NotI/XhoI opened pCAGGS-based expression plasmid.
pCAGGS-E-hPELI1-PM is useful for highly efficient expression of E-tagged hPELI1 with a mutated RING-domain 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 the human PELI1-PM cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF302505.1, except for the double point mutation H369S/C371S as well as for the nucleotides at positions 72, 149 and 243 of the coding sequence which do not change the amino acid sequence, and at position 149 which results in a N97T change compared to the EMBL record. The hPELI1-PM insert was sequenced by the depositor.
Other name of the plasmid is pCAGGS-E-hPeli-1PM.
|EMBL Accession number:||AF302505.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||16/04/2009|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: Alw44I/SacII, HincII, NcoI/XmnI, NotI/XhoI and SspI.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by Dr R. Schauvliege(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Schauvliege et al., FEBS Lett. 580 (2006), 4697-4702 [PMID: 16884718]
|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-E-hPELI1-PM (LMBP 5192) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Schauvliege et al., 2006.|
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.