棉蚜para钠通道cDNA和基因组DNA片段的克隆和序列分析

CLONING AND SEQUENCE ANALYSIS OF cDNA AND GENOMIC DNA FRAGMENTS ENCODING PARA SODIUM CHANNEL FROM APHIS GOSSYPII GLOVER

采用降落 PCR技术 ,根据昆虫 para型钠通道 区 S4至 S6及 与 连接区的保守区域设计简并引物 ,成功地克隆了棉蚜 para型钠离子通道 c DNA和基因组 DNA片段 (Gene Bank登录号分别为 :AF4 114 5 5、AF4 475 74 ) ,其中 c DNA片段为 6 5 9bp,编码 2 2 0个氨基酸。利用 c DNA片段设计特异性引物克隆获的基因组DNA片段为 132 8bp,共有 2个内含子。Blast序列分析表明 ,PCR扩增获得的棉蚜 para型钠离子通道 c DNA片段所编码的氨基酸与其他昆虫的 para型钠通道氨基酸具有很高的同源相似性 ,与马铃薯甲虫、德国小蠊、果蝇和家蝇的同源性分别为 6 5 %、6 3%、6 3%、5 9%。棉蚜 para型钠离子通道 c DNA和基因组 DNA片段的克隆对于农药分子设计与害虫抗性机制 ,特别是靶标抗性分子监测具有重要意义。

Genomic DNA and cDNA fragments encoding para sodium channel were cloned from cotton aphid, Aphis gossypii Glover using Touch down PCR technique with degenerate primer. The amplified cDNA fragment is of 659bp, which demonstrate high identity with para sodium channel gene amplified from other insects through Blast sequence analysis. Genomic DNA is of 1328bp, including 2 introns. Cloning of cDNA fragments from Aphis gossypii bear great significance on the pesticides molecular design and pest resistance research., especially pest resistance molecular monitoring.