The construction of shuttle vectors of Brevibacillus brevis-Escherichia coli.
- Author:
Qing-Zhong PENG
1
;
Wei-Cai ZHANG
;
Hou-Chu ZHU
Author Information
1. Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Bacillus;
drug effects;
genetics;
Cloning, Molecular;
Drug Resistance, Microbial;
genetics;
Erythromycin;
pharmacology;
Escherichia coli;
drug effects;
genetics;
Genetic Vectors;
genetics;
Plasmids;
genetics;
Recombinant Fusion Proteins;
genetics;
metabolism;
alpha-Amylases;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2002;18(4):438-441
- CountryChina
- Language:Chinese
-
Abstract:
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.
