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茶叶科学 ›› 2010, Vol. 30 ›› Issue (1): 37-44.doi: 10.13305/j.cnki.jts.2010.01.006

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茶尺蠖普通气味结合蛋白(GOBP)的基因克隆与序列分析

陈华才1, 刘军1, 张晓燕1, 殷坤山2   

  1. 1. 中国计量学院,浙江 杭州 310018;
    2. 中国农业科学院茶叶研究所,浙江 杭州310008
  • 收稿日期:2008-12-22 修回日期:2009-08-13 发布日期:2019-09-10
  • 作者简介:陈华才(1968— ),男,湖北浠水人,研究员,博士,研究方向为昆虫化学生态与分子生物学。huacaichen@cjlu.edu.cn
  • 基金资助:
    浙江省自然科学基金(Y305130)

Cloning and Sequence Analysis of EoblGOBP in the Antennal of Ectropis obliqua hypulina

CHEN Hua-cai1, LIU Jun1, ZHANG Xiao-yan1, YIN Kun-shan2   

  1. 1. China Jiliang University, Hangzhou 310018, China;
    2. Tea Research Institute of Chinese Academy of Agricultural Sciences, Hangzhou 310008, China
  • Received:2008-12-22 Revised:2009-08-13 Published:2019-09-10

摘要: 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%。

关键词: 茶尺蠖, 普通气味结合蛋白(GOBP), 全长序列, 基因克隆, 序列分析

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%.

Key words: Ectropis oblique, general odorant binding protein, full-length sequence, gene cloning, sequence analysis

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