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Cloning and sequencing of cDNA encoding general odorant binding protein Ⅱ in the antenna of Helicoverpa assulta ( Guenée ) and its expression in Escherichia coli
Author(s): 
Pages: 18-23
Year: Issue:  1
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  烟实夜蛾普通气味结合蛋白触角基因克隆原核表达;
Abstract: 利用RT-PCR技术扩增了编码烟实夜蛾Heltcoverpa assulta雌、雄虫触角普通气味结合蛋白Ⅱ的cDNA片段,将其克隆至pGEM-T Easy载体,获得了普通气味结合蛋白Ⅱ基因成熟蛋白阅读框序列.将该基因重组到表达型质粒pET-30a(+)中,并转化入原核细胞中表达.序列测定结果表明,烟实夜蛾触角普通气味结合蛋白基因的成熟蛋白阅读框全长489 bp,编码162个氨基酸残基,预测分子量和等电点分别为18.2 kD和5.35.推导的氨基酸序列与已报道的10种昆虫普通气味结合蛋白Ⅱ高度同源(73%~98%),并具有气味结合蛋白的典型特征.SDS-PAGE和Western印迹分析表明,经IPTG诱导,普通气味结合蛋白Ⅱ基因能在大肠杆菌BL21(DE3)中表达,电泳检测到一条约23 kD大小的外源蛋白,与预测的融合蛋白分子量大小相应.
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