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利用DNAman设计简并引物,通过反转录一多聚酶链式反应(RT-PCR)克隆小菜蛾Plutella xylostella(L.)抗性、敏感种群中可能与苏云金芽孢杆菌(Bt)抗性相关的碱性磷酸酶基因。琼脂糖电泳结果显示扩增条带与预期片段长度一致,阳性克隆经测序获得了403bp的基因片断。经blast比较得出所克隆基因属于碱性磷酸酶基因。与敏感种群相比,抗性种群中该基因片段有18个碱基发生变化,有1个氨基酸发生替换(缬氨酸转变为苏氨酸)。研究结果对于进一步研究小菜蛾全基因结构、功能有重要的意义。
Degenerate primers were designed by using DNAman and the basicity related to Bacillus thuringiensis (Bt) resistance in susceptible and susceptible populations of the diamondback moth Plutella xylostella (L.) was cloned by reverse transcription-polymerase chain reaction (RT-PCR) Phosphatase gene. The results of agarose gel electrophoresis showed that the amplified fragment was consistent with the expected fragment length, and the positive clone was sequenced to obtain 403 bp gene fragment. The blast result shows that the cloned gene belongs to the alkaline phosphatase gene. Compared with the susceptible population, there were 18 bases in the resistant population and one amino acid was substituted (valine to threonine). The results are of great significance for further study of the whole genome structure and function of Plutella xylostella.