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%。
Abstract
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%.
关键词
茶尺蠖 /
基因克隆 /
普通气味结合蛋白（GOBP） /
全长序列 /
序列分析
Key words
Ectropis oblique /
full-length sequence /
gene cloning /
general odorant binding protein /
sequence analysis
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] 孙晓玲, 陈宗懋. 基于化学生态学构建茶园害虫无公害防治技术体系[J]. 茶叶科学, 2009, 29(1): 136-143.
[2] 熊兴平. 茶尺蠖防治技术研究进展及展望[J]. 中国茶叶, 2003, 25(13): 15-17.
[3] 王桂荣, 郭予元, 吴孔明. 昆虫触角气味结合蛋白的研究进展[J]. 昆虫学报, 2002, 45(1): 131-137.
[4] Vogt R G, Riddiford L M.Pheromone binding and inactivation by moth antennae[J]. Nature, 1981(293): 161-163.
[5] Pelosi P, Maida R.Odorant-binding proteins in insects[J]. Comp Biochem Insect Biochemistry and Molecular Biology, 1995, 111(5): 503-514.
[6] 朱彬彬, 姜勇, 牛长缨, 等. 家蝇气味结合蛋白基因cDNA片段的克隆与序列分析[J]. 昆虫学报, 2005, 48(5): 804-809.
[7] 李红亮, 聂文敏, 高其康, 等. 中华蜜蜂气味结合蛋白ASP2 cDNA的克隆及原核表达[J]. 中国农业科学, 2008, 41(3): 933-938.
[8] Gui L Y, Chen Z M, Liu S S.Effect of exogenous methyl jasmonate-induced tea volatiles on host-selection behavior of insects[J]. 茶叶科学, 2004, 24(3): 166-171.
[9] Scaloni A, Monti M, Angeli S.Structural analysis and disulfide bridge pairing of two odorant binding proteins from Bombyx mori[J]. Biochemistry Physiology Research Communication, 1999(266): 386-391.
[10] Leal W S, Nikonoval L, Peng G.Disulfide structure of the pheromone binding protein from the silkworm moth, Bombyx mori[J]. Federation of European Biochemical Societies, 1999(464): 85-90.
[11] Damberger F, Nikonova L, Horst R.NMR characterization of a pH-dependent equilibrium between two folded solution conformations of the pheromone-binding protein from Bombyx mori[J]. Protein Science, 2000, 9(5): 1038-1041.
[12] 赵新成, 阎云花, 王琛柱. 雄性棉铃虫和烟青虫对雌性信息素的触角电生理反应[J]. 动物学报, 2003, 49(6): 795-799.
[13] 刘晓光, 安世恒, 罗梅浩, 等. 烟实夜蛾信息素结合蛋白3 cDNA 的克隆、序列分析与原核表达[J]. 昆虫学报, 2006, 49(5): 733-739.
PDF(329 KB)
浙公网安备 33019902000101号