以触角总RNA为模板,根据同源性设计简并性引物,采用RT-PCR扩增结合RACE技术克隆获得了茶尺蠖（Ectropis obliqua）普通气味结合蛋白GOBP1和GOBP2基因的cDNA全长序列。分别命名为EoblGOBP1和EoblGOBP2。EoblGOBP1的cDNA全长为1 528 bp,开放阅读框501 bp,编码166个氨基酸残基,预测分子量Mw=18 835.63 D,等电点pI=5.45,cDNA和氨基酸序列在NCBI/GenBank的登录号分别为FJ156732和ACN29680.1。EoblGOBP2的cDNA全长为1 315 bp,开放阅读框405 bp,编码160个氨基酸残基,预测分子量Mw=18 002.75 D,等电点pI=5.32。cDNA和氨基酸序列在NCBI/GenBank的登录号分别为FJ156733和ACN29681.1。两个气味结合蛋白的氨基酸序列中都含有6个保守的半胱氨酸位点,具有气味结合蛋白的典型特征,与已报道的鳞翅目昆虫气味结合蛋白的同源性为分别62%~82%和76%~88%。
The full-length sequences of two general odorant binding protein (GOBP) genes in Ectropis obliqua (named EoblGOBP1 and EoblGOBP2) were cloned with degenerated digonudeotide primers by RT-PCR and the rapid amplification of cDNA ends (RACE) using the total RNA isolated from the antenna of Ectropis obliqua as template. The length of EoblGOBP1 was 1 528 bp with a 501bp ORF which encoded a peptide of 166 amino acid residues. The predicted Mw and pI of the peptide were 18 835.63 D and 5.45 respectively. The NCBI/GenBank accession numbers of cDNA and amino acid of EoblGOBP1were FJ156732 and ACN29680.1, respectively. The length of EoblGOBP2 was 1 315 bp with a 405 bp ORF which encoded a peptide of 160 amino acid residues. The predicted Mw and pI of the peptide were 18 002.75 D and 5.32 respectively. The GenBank accession numbers of cDNA and amino acid of EoblGOBP2 were FJ 156733 and ACN29681.1, respectively. The amino acid sequences from the two gene sequences had 6 typical conservative Cys. Blast searching in NCBI showed that the amino acid sequences were highly similar to that of other GOBPs from Lepidoptera with sequence identities of 62%~82% and 76%~88%.
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