Last data update: 24 January 2024 16:39 CET
Plasmid name: pPLc28BHTNFm28Fs (LMBP 1799)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p1799.gb |
| Sequence analysis results Genecorner: |
- |
| 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: |
Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp) |
| Terminator: | - |
| 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. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| 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 [DOI: 10.1038/nbt1183-784] |
| 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] |
| Helper plasmid: | pcI857 |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pPLc28BHTNFm28Fs (LMBP 1799) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Simon et al., 1983. |
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.