Last data update: 21 October 2020 04:26 CEST
Plasmid name: pCAGGSE6L (LMBP 3547)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) cDNA; light chain (E6L)
|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; pSV51E6L1|
|Further information:||The plasmid was constructed by ligating the XbaI fragment (filled in with Klenow DNA polymerase) from pSV51E6L1, containing a cDNA copy of the light chain of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ), to the MscI vector fragment from pCAGGS.
pCAGGSE6L is useful for efficient expression of the light chain of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) under the control of the chicken β-actin/rabbit β-globin hybrid promoter (AG) and the human CMV-IE enhancer in various mammalian cells.
In combination with an E6H (heavy chain of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ)) expression vector (e.g. pCAGGSSE6H), this plasmid can be used to produce a hPLAP (human placental alkaline phosphatase)-specific mouse antibody.
In combination with an expression vector for a heavy chain mouse immunotoxin (e.g. pUHDE6Hf2ETA) or a mouse::human immunotoxin (e.g. pUHDE6Hy3f2ANG), this plasmid can be used to produce a hPLAP-specific mouse or mouse::human chimeric immunotoxin respectively.
The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon.
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 LT727350.1.
|EMBL Accession number:||LT727350.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||31/08/2000|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, BamHI/XmnI, BglII/XhoI, SacII, SacII/SspI, SspI and XmnI.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Contreras(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061(λ)|
|Host reference:||Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
|Related host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGSE6L (LMBP 3547) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Contreras .|
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.