摘要
本文以质粒pE194为载体亚克隆B.licheniformis热稳定α-淀粉酶基因,构建成重组质粒pNW102,通过噬菌体PBS1将它转导进入中温α-淀粉酶生产菌B.subtilis BF7658。B.subtilis BF7658(pNW102)经过长时间非许可温度处理,筛选得到2株热稳定α-淀粉酶稳定性表达的工程菌株。酶学分析显示同源重组具有热点,2株重组菌株B.subtilis BFNW产生的热稳定α-淀粉酶符合B.licheniformis产生的淀粉酶特性。
The plasmid pE194 as a vector is used to subclone the thermostable a-amylase gene of ft licheniformts and construct recombinant plasmid pNW102. The plasmid pNW102 was transduced into B. subilis BF 7658 by bacteriophage PBS1. The B. subtilis BF7658(pNW102) strain, was treated at non-permissive temperature for a long time, and obtained many recombinant strains. This is the homologous recombination results between a-amylase genes of pNW102 and BF7658. The homologous recombination occurs at any site in a-amylase gene but there are some heat-spots. We have already screened 2 strains from the recombination strains which can stably produce the thermostable a-amylase in B. subtilis BF7658. The analysis of enzymology shows that the characters of the thermpstable a-amylase produced from the recombinant! strains are the same with that of B. licheniformis.
基金
国家自然科学基金
关键词
热稳定
Α-淀粉酶
工程菌
Subcloning, Transduction, Homologous recombination, Thermostable aamylase
