Anthony N, Rocheleau T, Mocelin G, Lee H J, ffrench-Constant R
Department of Neuroscience, University of Wisconsin at Madison 53706, USA.
FEBS Lett. 1995 Jul 24;368(3):461-5. doi: 10.1016/0014-5793(95)00711-h.
A degenerate PCR strategy was used to isolate a fragment of the acetylcholinesterase gene (Ace) homolog from Aedes aegypti and screen for a cDNA clone containing the complete open reading frame of the gene. The predicted amino acid sequence of the Aedes gene shares 64% identity with Ace from Drosophila and 87% identity with the acetylcholinesterase gene from another mosquito species Anopheles stephensi. High levels of expression of the Aedes gene were achieved by infection of Sf21 cells with a recombinant baculovirus containing the Aedes Ace cDNA. The catalytic properties and sensitivity of the recombinant enzyme to insecticide inhibition are described and discussed in relation to the role of insensitive AChE in conferring resistance to organophosphorus and carbamate insecticides.
采用简并PCR策略从埃及伊蚊中分离出乙酰胆碱酯酶基因(Ace)同源物的一个片段,并筛选出包含该基因完整开放阅读框的cDNA克隆。埃及伊蚊基因的预测氨基酸序列与果蝇的Ace有64%的同一性,与另一种蚊子斯氏按蚊的乙酰胆碱酯酶基因有87%的同一性。通过用含有埃及伊蚊Ace cDNA的重组杆状病毒感染Sf21细胞,实现了该基因的高水平表达。针对不敏感乙酰胆碱酯酶在赋予对有机磷和氨基甲酸酯类杀虫剂抗性中的作用,描述并讨论了重组酶的催化特性和对杀虫剂抑制的敏感性。