小菜蛾碱性磷酸酶基因cDNA片段的克隆及其序列分析
Cloning and sequence analysis of alkaline phosphatase gene in the diamondback moth, Plutella xylostella.
刘洁1，2** ,文礼章1 ,王少丽2 ， ,吴青君2 ，张友军2***
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作者单位：湖南农业大学生物安全科学技术学院长沙410128； 2.中国农业科学院蔬菜花卉研究所北京100081
中文关键词：小菜蛾， 碱性磷酸酶酶基因，克隆，序列分析
英文关键词：Plutella xylostella, alkaline phosphatase, cloning, sequence analysis
中文摘要：
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403 bp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。
英文摘要： Alkaline phosphatase genes were cloned from the resistant and susceptible strains of Plutella xylostella(L.) through RT-PCR technique using degenerate primers and sequenced. The cDNA length was 403 bp and the sequence was confirmed to be the alkaline phosphatase gene by blast alignment(BLAST). The nucleotide similarity of the resistant and susceptible strains was 99.5% and the amino acids identity was 99%. Compared with the susceptible strain, 18 nucleotides were changed in the resistant strain, resulting in one amino acid changed (V to T). These results are helpful for further study of the full sequence and function of alkaline phosphatase gene in P. xylostella.