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Cloning and sequencing of cDNA encoding a chemosensory protein in the antenna of Helicoverpa assulta and its expression in Escherichia coli
Author(s): 
Pages: 740-746
Year: Issue:  5
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  烟实夜蛾化学感受蛋白基因克隆测序原核表达;
Abstract: 利用RT-PCR技术扩增了编码烟实夜蛾Helicoverpa assulta触角化学感受蛋白(chemosensory protein)的全长cDNA.克隆和测序结果表明,烟实夜蛾化学感受蛋白基因核苷酸序列全长384bp(GenBank序列号:DQ285667),编码127个氨基酸残基,预测N-末端包含16个氨基酸组成的信号肽序列,因此估测其成熟蛋白分子量为12.97kD,等电点为5.32.将该基因重组到表达载体pGEX-4T2中,并转入原核细胞中进行表达.SDS-PAGE和Westem印迹分析表明,经IPTG诱导后,烟实夜蛾化学感受蛋白基因能在大肠杆菌BL21中表达,电泳检测到一条约39 kD的外源蛋白,与预测的融合蛋白分子量大小相符.
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