Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLT10igaT (LMBP 3241)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease (iga)|
|Promoter:||Phage λ major leftward promoter (λ PL)
Phage T7 gene 10 promoter (T7g10)
|Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)|
|Terminator:||Phage T7 gene 10 terminator (T7g10)|
|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:||pLT10T; pMc514igam1|
|Further information:||The plasmid was constructed via a three-point-ligation of the following fragments: 1) the BsmI (blunted with T4 DNA polymerase) - BglII fragment of pMc514igam1, encoding the N-terminal part of the IgA protease gene; 2) the BglII-HindIII fragment of pMc514igam1, encoding the C-terminal part of the IgA protease gene; 3) the ApaI (blunted with T4 DNA polymerase) - HindIII vector fragment of pLT10T.
This is an expression plasmid for the IgA1 protease precursor under control of the PL or the T7 promoter.
For T7 driven expression use a strain containing a controllable T7 RNA polymerase gene, as well as a cI function: preferably a pT7POL plasmid (Mertens et al., Biotechnology 13 (1995), 175-179) or e.g. BL21(DE3)[pcI857]. Proceed as follows: first transform auxiliary plasmid, make competent cells again and then transform the expression plasmid.
To reconstruct the nucleotide sequence of this plasmid, the sequence of the IgA1 protease gene from Neisseria gonorrhoeae MS11 corresponds with the EMBL Nucleotide Sequence Database (Release 36; Accession number X04835). However, the cloned IgA1 protease gene was derived from Neisseria gonorrhoeae NCTC8375 and differs in several nucleotides. As such, there is uncertainty as to the cutting frequency of the restriction enzymes indicated.
Other name of the plasmid is pLT10tIgAp.
|EMBL Accession number:||-|
|Latest sequence update:||30/06/1995|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by I. Poelaert(1) (2) and 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:||PhD thesis Irmgard Poelaert (1995)
|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.