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

Cloning and knockout vector construction of a NAD(P)H: quinone oxidoreductase gene in Bacillus thuringiensis

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pRN5101DKU,为后续Bt8010菌株NAD(P)H:醌氧化还原酶基因的敲除做准备.

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.The 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.