Merryweather A T, Crampton J M, Townson H
Wolfson Unit of Molecular Genetics, Liverpool School of Tropical Medicine, U.K.
Biochem J. 1990 Feb 15;266(1):83-90. doi: 10.1042/bj2660083.
Organophosphate-resistant and -susceptible strains of Culex quinquefasciatus (mosquito) have been compared on the basis of their esterase activities. The homozygous resistant strain (Dar) shows two highly active esterases after starch-gel electrophoresis, of Rm 0.2 and 0.4, which are absent from susceptible strains (Apo, Mon), and which previous selection studies have shown to be inseparable from organophosphate resistance. After SDS/polyacrylamide-gel electrophoresis and silver staining of total C. quinquefasciatus proteins, a 62 kDa band is observed in strain Dar at high concentrations, and in susceptible strains in trace amounts. After Western blotting, this 62 kDa protein is recognized by antisera raised against the two esterases eluted from starch gels. After chromatofocusing of Dar proteins, the 62 kDa protein is seen to be associated with esterase activity, and of a similar pI to that observed for esterases after isoelectric focusing. Post-translational modification is not required for recognition of the 62 kDa putative esterase, since the protein is immunoprecipitated by the anti-esterase serum from products of translation of Dar mRNA in vitro.
基于酯酶活性，对致倦库蚊（蚊子）的有机磷抗性和敏感品系进行了比较。纯合抗性品系（Dar）在淀粉凝胶电泳后显示出两种高活性酯酶，迁移率（Rm）分别为0.2和0.4，而敏感品系（Apo、Mon）中不存在这两种酯酶，并且先前的选择研究表明它们与有机磷抗性不可分割。对致倦库蚊总蛋白进行SDS/聚丙烯酰胺凝胶电泳和银染后，在高浓度的Dar品系中观察到一条62 kDa的条带，在敏感品系中则为痕量。经过蛋白质印迹法后，这条62 kDa的蛋白质能被针对从淀粉凝胶上洗脱的两种酯酶制备的抗血清识别。对Dar品系的蛋白质进行色谱聚焦后，发现这条62 kDa的蛋白质与酯酶活性相关，其等电点与等电聚焦后观察到的酯酶相似。对这条62 kDa的假定酯酶的识别不需要翻译后修饰，因为该蛋白质能被抗酯酶血清从体外翻译的Dar品系mRNA产物中免疫沉淀下来。
