军事医学2011，Vol.35Issue(10)：754-757,799,5.
应用Red重组技术构建四环素抗性多基因串联表达载体
Construction of a tetracycline resistance multi-gene coexpression vector using Red recombineering
摘要
Abstract
Objective To construct a tetracycline resistance multi-gene coexpression vector. Methods Tetracycline resistance gene with homologous primers was cloned by overlap extension PCR before the resistance gene sequences in multi-gene coexpression vector pET-m28a( +) were replaced by tetracycline resistance gene in DH5α via Red-mediated recombination system pKD46. Positive recombinants were selected under tetracycline pressure. Using isocaudarners,sfp and accA1 were combined in pET-mt28a( +), and the expression of genes was identified. Results Positive recombinants with pET-m28a( +) and tetracycline resistance were selected. Restructuring plasmid pET/sfp/accAl-mt28a( +)was constructed, and the two genes could co-express in one vector. Conclusion We successfully construct a tetracycline resistance multi-gene coexpression vector pET-mt28a( +), contributing to the reconstruction of erythromycin synthesis pathways in E. Coli.关键词
四环素抗药性/多基因串联表达载体/Red/重组/质粒/红霉素Key words
tetracycline resistance/ multi-gene coexpression vector/ Red recombineering/ plasmids/ erythromycin分类
生物科学引用本文复制引用
何彰华,张乐之,李德彬,闫兴,赵志虎,师明磊,王洋,叶丙雨,黄芬,张彦,范秋声,王东,肖丽霞..应用Red重组技术构建四环素抗性多基因串联表达载体[J].军事医学,2011,35(10):754-757,799,5.基金项目
国家重大新药创制关键技术平台(2008ZXJ09007-01) （2008ZXJ09007-01）
