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Cloning and overexpression of transferrin gene from cypermethrin-resistant Culex pipiens pallens

机译:抗氯氰菊酯的淡色库蚊的转铁蛋白基因的克隆和过表达

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摘要

The complete sequence of transferrin has been cloned from cypermethrin-resistant strain of Culex pipiens pallens(Cr-C strain). Quantitative reverse transcription PCR analysis indicated that the transferrin transcription level was 12.6 times higher in Cr-C strain than in susceptible strain at fourth instar larvae. The transferrin expression was also found to be consistently higher throughout the life cycle of Cr-C strain. A protein of predicted size 90.8 kDa has been detected by Western blotting in transferrin-transfected mosquito C6/36 cells. These transferrin-transfected cells also showed enhanced cypermethrin resistance compared to null-transfected or plasmid vector-transfected cells as determined by methyl tritiated thymidine( 3H-TdR) incorporation. These results indicate that transferrin is expressed at higher levels in Cr-C strain and may confer some insecticide resistance in C. pipiens pallens.
机译:已从淡色库蚊对氯氰菊酯抗性菌株(Cr-C菌株)克隆了转铁蛋白的完整序列。定量逆转录PCR分析表明,在四龄幼虫中,Cr-C株的转铁蛋白转录水平比易感株高12.6倍。还发现在Cr-C菌株的整个生命周期中,转铁蛋白的表达始终较高。通过Western印迹在转铁蛋白转染的蚊子C6 / 36细胞中检测到了预测大小为90.8 kDa的蛋白质。这些转铁蛋白转染的细胞与通过甲基化胸苷(3H-TdR)掺入确定的空转染或质粒载体转染的细胞相比,还显示出增强的氯氰菊酯抗性。这些结果表明,转铁蛋白在Cr-C菌株中以较高水平表达,并可能在淡色梭菌中赋予一定的杀虫剂抗性。

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