Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLc28BHTNFm28Fs (LMBP 1799)
|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); mutated 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:||pPLc28BHTNF; pMaT4HTNFTm28F|
|Further information:||The plasmid was constructed by inserting the AvaI-BstEII fragment of pMaT4HTNFTm28F, containing the
mutated mature hTNF sequence, into the AvaI-BstEII opened pPLc28BHTNF plasmid.
The plasmid contains the amp signal sequence, fused in frame to the mutated mature hTNF sequence.
The first 7 amino acids of the mature hTNFm28F are identical to wild type, but are encoded by the synthetic linker sequence.
Strains, containing pPLc28BHTNFm28F, secrete mature hTNFm28F into the periplasmic space, in a temperature dependent mode.
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.
|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 R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Simon et al., Biotechnology 1 (1983), 784-791
|Related plasmid reference:||Van Ostade et al., FEBS Lett. 238 (1988), 347-352 [PMID: 3049162]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 K514[pcI857]|
|Host reference:||Zabeau et al., EMBO J. 1 (1982), 1217-1224 [PMID: 6327257]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 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.