摘要
目的 从白纹伊蚊中分离、克隆及鉴定乙酰胆碱酯酶 (ACh E)基因片段。 方法 根据已知的果蝇和斯氏按蚊 ACh E氨基酸序列保守区域设计简并引物 ,对白纹伊蚊 ACh E基因片段进行扩增 ,将凝胶回收的 PCR产物经与 T-载体质粒连接进行 T/ A克隆 ,用α互补筛选法筛选重组质粒克隆 ,用碱裂解法提取重组质粒 DNA并对重组质粒进行酶切鉴定和 PCR鉴定。 结果与结论 获得与设计相符的白纹伊蚊 ACh E基因片段
Objective To isolate, clone and identify the acetylcholinesterase (AChE) fragment from the mosquito, Aedes albopictus, in relation to exploring mechanism of insecticide resistance. \ Methods\ The genome DNA extracted from the mosquito was used for degenerate polymerase chain reaction (PCR) and the two pairs of oligonucleotides encoding the highly conserved protein sequences were used as primers. The reaction products were cloned to T\|vector and transfected into E\^coli JM 109. The replicative form DNA of recombinant vector extracted from E\^coli JM 109 through alkalilysis was identified by the methods of digestion with EcoRⅠand SalⅠ and PCR. \ Results\ The products of degenerate primers polymerase chain reaction were obtained and the identified clone belongs to the AChE fragment of the mosquito.\ Conclusion\ The clone was identified as the AChE fragment of Aedes albopictus.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2001年第4期209-212,共4页
Chinese Journal of Parasitology and Parasitic Diseases
关键词
白纹伊蚊
乙酰胆碱酯酶
抗药性
克隆
鉴定
Aedes albopictus, acetylcholinesterase, insecticide resistance, clone.
