Last data update: 28 November 2020 04:24 CET
Plasmid name: pMa58iga (LMBP 3237)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease (iga)|
|Ribosome binding site (RBS) of the Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease (igaT)|
|Terminator:||Phage fd terminator|
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was constructed as follows: 1) Via the PCR reaction, a unique SmaI restriction site was created in the 5' UTR region of the Neisseria gonorrhoeae NCTC8375 iga gene, and a unique XbaI site was created in the 3' UTR region. The synthetic linkers used for this PCR reaction were 5'-CCCAACCCGGGTAAAATGCAAAACATTATCGC-3' and 5'-GGCATTGAGCTCTAGATTTGTTAGTCTG-3' respectively; 2) The IgA1 protease gene was then isolated as a 4.7 kb SmaI-XbaI fragment and ligated to the SmaI-XbaI vector fragment of pMa58.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
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 pMa58IgAp.
|EMBL Accession number:||-|
|Latest sequence update:||31/07/1995|
|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|
|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 + 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.