关键词: 淡色库蚊, 杀虫剂抗性, 基因克隆, 抑制性差减杂交, 基因芯片, 逆Northern

Abstract: 　Objective To obtain deltamethrin-resistance or susceptibility associated genes of Culex pipiens pallens. \ Methods\ The differentially expressed genes were obtained by suppression subtractive hybridization (SSH), and identified by cDNA microarray and reverse Northern blotting.\ Results\ 523 and 286 clones were selected respectively in the two directional SSH. 155 and 42 genes were respectively expressed 2-3 and >3 times higher in the insecticide-resistant strain than in the susceptible strain; 15 and 9 genes were respectively expressed 2-3 and >3 times higher in the susceptible strain than in the resistant strain. There were 2 genes only expressed in the insecticide-resistant strain. 51 three times differentially expressed clones and 2 specially expressed clones were sequenced. 44 sequences were obtained which belong to 13 new genes. There were 8 over-expressed genes in resistant strain, 7 of which were similar respectively to mitochondrion rRNA gene, 60S ribosomal protein gene, 40S ribosomal protein S4 gene, trypsin gene, chymotrypsin A gene, opsin gene, and 16S ribosomal RNA gene. There were 5 over-expressed genes in susceptible strain, 2 of them being similar with 40S ribosomal protein S29 gene and myosin regulatory light chain 2 gene. In addition, 2 genes specially expressed in resistant strain were similar respectively to glycogen branching enzyme gene and ribosomal protein 46 gene.\ Conclusion\ The differentially expressed genes may be associated with deltamethrin-resistance or susceptibility of Culex pipiens pallens.

Key words: Culex pipiens pallens, insecticide resistance, gene cloning, suppression subtractive hybridization, cDNA microarray, reverse Northern