Last data update: 28 November 2020 04:24 CET
Plasmid name: pMG6 (LMBP 8457)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Bacillus subtilis amylase gene (amyE); 5' UTR and 3' UTR|
|Promoter:||Escherichia coli lac operon promoter|
|Selection marker:||Ampicillin (amp)
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli
Bacillus sp.; integrative
|Further information:||The plasmid was constructed by removing the PstI and HindIII sites downstream of the 3' UTR of amyE in the pMG1 vector. The construction of the pMG1 vector based on pUC18 and pUC19 is described in Gimpel et al. (2012).
This plasmid is a modular E. coli–Bacillus shuttle vector, applicable for gene knockout in B. subtilis via chromosomal integration.
For this purpose the 5' UTR and 3' UTR sequences of amyE have to be replaced with the 5' UTR and 3' UTR of the gene to be knocked out. They can be replaced as an AatII/XbaI fragment and SalI/NotI fragment respectively.
Furthermore, the modular architecture of the pMG plasmid family enables an easy exchange of all features like antibiotic resistance genes (EcoRI/SalI), promoters (EcoRI/PstI), 5′ UTR (PstI/HindIII) and tags (HindIII/Acc65I) leading to a significant increase of applicabilities.
The nucleotide sequence of the B. subtilis fragments correspond with the EMBL Nucleotide Sequence Database accession number AP012496.1.
|EMBL Accession number:||AP012496.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||06/06/2013|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: EcoRI/SalI, HaeII, HincII and NotI.|
|History of deposit:||This plasmid was deposited by Dr. M. Gimpel(1) and Prof. Dr S. Brantl(1).
(1) AG Bakteriengenetik, FSU Jena, Jena, Germany
|Plasmid reference:||Gimpel et al., J. Microbiol. Methods 91 (2012), 312-317 [PMID: 22982324] [DOI: 10.1016/j.mimet.2012.09.003]
|Restricted distribution:||- BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + chloramphenicol (34 μg/ml)*|
|Cultivation remark:||*: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin.|
|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.