苏云金芽孢杆菌NAD(P)H:醌氧化还原酶基因的克隆和敲除载体的构建

摘要

Bacillus thuringiensis subsp. Kurstaki isolate 8010 was selected to obtain the upstream and downstream sequences of NAD(P)H: quinone oxidoreductase gene by PCR technology. Trie pDG780 vector (containing the kanamycin resistance gene) was used to construct a recombination plasmid pRN5101DKU which contained the upstream and downstream sequences of NAD(P)H; quinone oxidoreductase , and the kanamycin resistance gene by PCR and RE treatments. It was provided for further study on gene knockout.%以苏云金芽孢杆菌库斯塔克亚种(Bacillus thuringiensis subsp.kurstaki)8010为研究材料,通过PCR技术获得Bt 8010菌株NAD(P)H:醌氧化还原酶基因上下游2个片段,以含有卡那霉素抗性基因的pDG780质粒为中间载体,通过酶切、连接和PCR,构建含待敲除NAD(P)H:醌氧化还原酶基因上下游片段、卡那霉素抗性基因的重组质粒pRN5 101DKU,为后续Bt8010菌株NAD(P)H:醌氧化还原酶基因的敲除做准备.