[The construction of shuttle vectors of Brevibacillus brevis-Escherichia coli].
The 5' region of the cell wall protein(CWP) gene containing multiple tandem promoters and the signal peptide-coding sequence was isolated by PCR from Br. brevis 50, and used to construct the shuttle vector pBKE50, which included the replication origin of pUB110 and the erythromycin-resistance gene of pGK12. The alpha-amylase gene of Bacillus subtilis 168 was ligated to pBKE50, producing plasmid pBKE50/alpha-amy. After the resulting plasmid was introduced into Br. brevis 50, soluble and biologically active alpha-amylase was secreted directly into the culture medium. The expression level of alpha-amylase in the recombinant Br. brevis 50 was twice higher than that of the donor strain.
Peng
Qing-Zhong
QZ
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Zhang
Wei-Cai
WC
Zhu
Hou-Chu
HC
D001407:Bacillus / Q000187:drug effects / Q000235:genetics*; D003001:Cloning, Molecular; D004352:Drug Resistance, Microbial / Q000235:genetics; D004917:Erythromycin / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D004917:Erythromycin; D000516:alpha-Amylases