Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability.
A series of plasmid-based expression vectors have been constructed allowing stable intracellular expression of recombinant proteins in Bacillus subtilis strains. These expression vectors are based on the recently described Escherichia coli-B. subtilis shuttle vector pMTLBS72 which replicates as theta circles. Besides the weak constitutive promoter P(lepA), we inserted three different controllable promoters: P(gsiB) which can be induced by heat and acid shock, and by ethanol, P(xylA) and P(spac) which respond to the addition of xylose and IPTG, respectively. The versatility of these expression vectors was demonstrated by fusing their promoters to a reporter gene and by overexpression of the HtpG protein with three of them. All recombinant vectors exhibited full structural stability.
Nguyen
Hoang Duc
HD
Institute of Genetics, University of Bayreuth, D-95440 Bayreuth, Germany; Vietnam National University-Ho Chi Minh City, College of Natural Sciences, Faculty of Biology, Ho Chi Minh City, Viet Nam.
Nguyen
Quynh Anh
QA
Ferreira
Rita C
RC
Ferreira
Luis C S
LC
Tran
Linh Thuoc
LT
Schumann
Wolfgang
W
D001412:Bacillus subtilis / Q000235:genetics* / Q000254:growth & development; D004261:DNA Replication; D004269:DNA, Bacterial; D004270:DNA, Circular; D004926:Escherichia coli / Q000235:genetics; D000431:Ethanol / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005822:Genetic Vectors; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018869:Heat-Shock Response; D006358:Hot Temperature; D015151:Immunoblotting; D007544:Isopropyl Thiogalactoside / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D012769:Shock; D014994:Xylose / Q000378:metabolism; D001616:beta-Galactosidase / Q000032:analysis / Q000378:metabolism
D004269:DNA, Bacterial; D004270:DNA, Circular; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D000431:Ethanol; D014994:Xylose; D001616:beta-Galactosidase