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Cloning and sequencing of cDNA encoding pheromone binding protein 3 from the Helicoverpa assulta (Guenée) and its expression in Escherichia coli
Author(s): 
Pages: 733-739
Year: Issue:  5
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  烟实夜蛾信息素结合蛋白基因克隆原核表达;
Abstract: 利用RT-PCR技术从烟实夜蛾Helicoverpa assulta(Hass)雄虫触角中扩增得到了信息素结合蛋白3(Hass PBP3).克隆和测序结果表明,该基因核苷酸序列全长495bp,编码164个氨基酸残基,预测分子量18.5kD.并预测N-末端疏水区包含由22个氨基酸组成的信号肽.因此,成熟蛋白应包括142个氨基酸,预测分子量为16.1 kD,等电点为5.44.经氨基酸序列同源性分析发现,此序列与已知昆虫PBP3有较高的同源性,而且具有气味结合蛋白的典型特征.将该基因重组到表达载体pGEX-4T-2中进行原核表达.经IPTG诱导、SDS-PAGE分析和Western印迹检测,结果表明烟实夜蛾PBP3基因能在大肠杆菌BL21中表达,电泳检测到一条大约42kD的外源蛋白,与预测的融合蛋白分子量相符.
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