title: Construction of novel shuttle expression vectors for gene expression in Bacillus subtilis and Bacillus pumilus.
abstract: A native plasmid (pSU01) was detected by genome sequencing of Bacillus subtilis strain S1-4. Two pSU01-based shuttle expression vectors pSU02-AP and pSU03-AP were constructed enabling stable replication in B. subtilis WB600. These vectors contained the reporter gene aprE, encoding an alkaline protease from Bacillus pumilus BA06. The expression vector pSU03-AP only possessed the minimal replication elements (rep, SSO, DSO) and exhibited more stability on structure, suggesting that the rest of the genes in pSU01 (ORF1, ORF2, mob, hsp) were unessential for the structural stability of plasmid in B. subtilis. In addition, recombinant production of the alkaline protease was achieved more efficiently with pSU03-AP whose copy number was estimated to be more than 100 per chromosome. Furthermore, pSU03-AP could also be used to transform and replicate in B. pumilus BA06 under selective pressure. In conclusion, pSU03-AP is expected to be a useful tool for gene expression in Bacillus subtilis and B. pumilus.
authors.affiliation: Key Laboratory of Bio-Resource & Eco-Environment, Ministry of Education, Sichuan Key Laboratory of Molecular Biology & Biotechnology, College of Life Sciences, Sichuan University.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Shao
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Cao
authors.forename: Hongyan
authors.initials: H
authors.lastname: Zhao
authors.forename: Xuemei
authors.initials: X
authors.lastname: Tan
authors.forename: Hong
authors.initials: H
authors.lastname: Feng
mesh_terms: D001407:Bacillus / Q000235:genetics* / Q000254:growth & development; D001412:Bacillus subtilis / Q000235:genetics* / Q000254:growth & development; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D010450:Endopeptidases / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005822:Genetic Vectors; D010957:Plasmids
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010450:Endopeptidases; C087822:alkaline protease