Last data update: 26 September 2020 04:22 CEST
Plasmid name: pLysDNHIFNG (LMBP 2009)
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2009.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: | Human interferon γ cDNA (IFNG); mutated sequence Phage MS2 lysis protein gene |
| Promoter: | Phage λ major leftward promoter (λ PL) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp) |
| 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: | pPLc321HIFNGv; pLacDN3HIFNG; pT73MS2CL |
| Further information: | The plasmid was constructed by replacing the SpeI-XbaI (nucleotide position 479) fragment of pPLc321HIFNGv by a combination of the SpeI-EcoRI fragment of pLacDN3HIFNG and the EcoRI (nucleotide position 489) - XbaI (nucleotide position 54) fragment of pT73MS2C820. The mutated hIFNG gene (containing a unique SalI site at position 413 and an extra HhaI site at position 390 of the coding region; sequence represented at page 235) is fused to part of the MS2 coat protein gene in the reading frame of the lysis gene. This fusion protein contains 14 carboxy terminal missense amino acids. This is a detection system (DN) for nonsense mutants based on the MS2 lysis gene (Lys). Expression of the hIFNG/MS2-LYS fusion gene results in expression of the MS2 lysis gene, followed by lysis (the bacteria begin to lyse approx. 30 minutes after temperature induction from the PL-promoter). Other name of the plasmid is pDNBG. |
| EMBL Accession number: | - |
| Latest sequence update: | 13/03/1990 |
| 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 E. Sablon(1) and Prof. Dr E. Remaut(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| 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) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| 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.