Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLGN1HIL2 (LMBP 1247)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Human interleukin 2 cDNA (IL2, IL-2, TCGF, GeneID 3558); mature sequence|
|Promoter:||Phage λ major leftward promoter (λ PL)
Phage λ major rightward promoter (λ PR)
Phage λ promoter for repressor maintenance (λ PRM)
|Ribosome binding site (RBS) of the phage Mu ner protein gene|
|Selection marker:||Kanamycin (kan)|
|Replicon:||Broad-host-range plasmid RSF1010 vegetative replication origin; IncQ
Broad-host-range plasmid RSF1010 origin of transfer (oriT)
|Host range:||Escherichia coli
Gram-negative bacterial strains
|Parental clone:||pPLGN1; pPLc436HIL2|
|Further information:||The plasmid was constructed by cloning the fusion gene (signal sequence AMP-mature human IL2) as an EcoRI-BamHI(filled in with Klenow DNA polymerase) fragment from pPLc236BHIL2 between the unique EcoRI and PvuII sites of pPLGN1. The BamHI site is restored.
pPLGN1HIL2 is a broad-host-range plasmid, designed for expression of mature human IL2.
It can be used in all gram-negative bacterial species that support replication of the natural plasmid RSF1010.
The plasmid carries the cI857 gene encoding a temperature sensitive repressor. Induction of the PL promoter to synthesize mature human IL2 is obtained at 42°C (strains tested : E. coli, Serratia, Erwinia, Vibrio species).
Introduce the plasmid by electroporation, or through conjugation, using plasmid RK2/RP4 transfer functions (provided in trans by RK2/RP4, RK2/RP4-derived vectors, other replicons carrying RK2/RP4 transfer functions (e.g. pRK2073) or strains expressing the tra genes of RK2/RP4). Propagation temperatures may not exceed 34°C, preferentially grow at 28°C.
The nucleotide sequence of the mature human IL2 corresponds with the EMBL Nucleotide Sequence Database accession number X01586.1, except for the first codon of the mature sequence (GCA) that is replaced by GCG.
Other name of the plasmid is pMU1010.
|EMBL Accession number:||X01586.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||29/03/1995|
|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., J. Bacteriol. 169 (1987), 1899-1904 [PMID: 2952635]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|Cultivation medium:||LB-Lennox + 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.