Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLc28BHTNFs (LMBP 1708)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Ampicillin resistance gene (amp); signal sequence
Human tumor necrosis factor cDNA (TNF); mature sequence
|Promoter:||Escherichia coli β-lactamase promoter (amp)
Phage λ major leftward promoter (λ PL)
|Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp)|
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli; use strains with a cI function, cIts for PL controlled expression|
|Parental clone:||pPLc2820BA; pPLcmuHTNF1|
|Further information:||This plasmid was constructed as follows : to the ApaI site (blunted by T4 polymerase exonuclease treatment) of pPLc2820BA, an AvaI linker was ligated, and the AvaI-HindIII fragment of pPLcmuHTNF1 (containing the mature HTNF gene) was inserted.
The plasmid contains the AMP signal sequence, fused in frame with the mature human TNF coding sequence.
Strains, containing pPLc28BHTNF secrete mature human TNF into the periplasmic space.
The ampicillin resistance gene does not contain a PstI restriction site.
Expresssion in λ wt lysogen : constitutive synthesis under control of the amp promoter at any physiological temperature.
Expression in cIts : λ-PL induction at 42°C.
The first 7 amino acids of the mature human TNF are identical to wild type, but are encoded by a synthetic linker sequence (nucleotide sequence different from wild type human TNF)
Other name of the plasmid is pPLc2820BTNF.
|EMBL Accession number:||-|
|Latest sequence update:||29/08/1990|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Leemans et al., Gene 85 (1989), 99-108 [PMID: 2695405]
|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:||-|
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.