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GOBP1蛋白作为昆虫化学感受机制的重要蛋白,对于研究气味调控防治害虫有重要意义。采用RT-PCR技术以八字地老虎的触角总RNA为模板进行克隆得到1条1 260 bp的八字地老虎GOBP1基因片段(GenBank登录号为JX978464),编码161个氨基酸残基,预测分子量约为18.6 kD,等电点5.14。生物信息学在线预测分析表明:八字地老虎GOBP1核苷酸序列与黄地老虎Agrotis segetum(登录号为DQ838577.1)同源性最高,为97%,与其他昆虫的同源性达80%以上。构建了重组表达质粒pET21-b/Acni GOBP1原核表达后检测到18.6 kD的外源蛋白,Western bolt检测该蛋白是目的蛋白。结果为深入研究GOBP1基因以及通过气味调控防治害虫提供了基础资料。
GOBP1 protein as an important chemical mechanism of insects protein, for the study of odor control pest control is important. A total of 1 260 bp of the GOBP1 gene from GenBank (GenBank accession number JX978464) was cloned by RT-PCR using the total RNA of T. gigas as a template and encoded 161 amino acid residues with a predicted molecular mass of 18.6 kD, isoelectric point 5.14. Bioinformatics online predictive analysis showed that the nucleotide sequence of GOBP1 was 97% homologous to the yellow tiger Agrotis segetum (accession number DQ838577.1), and its homology with other insects was over 80% . The eukaryotic expression vector pET21-b / Acni GOBP1 was constructed to detect a foreign protein of 18.6 kD. The protein was identified by Western blot. The results provide the basic information for further study of GOBP1 gene and the control of pests through odor regulation.