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茶尺蠖脂肪酶合成基因EoL2的克隆和组织表达特征

杜琴, 辛肇军, 杨子威, 李喜旺, 陈宗懋, 孙晓玲   

  1. 中国农业科学院茶叶研究所/农业部茶树生物学与资源利用重点实验室, 杭州 310008
  • 收稿日期:2014-05-08 修回日期:1900-01-01 出版日期:2015-02-08 发布日期:2015-02-08
  • 通讯作者: 孙晓玲,博士,研究员,E-mail:xlsun1974@163.com

Clone and Expression Analysis of Lipase EoL2 Gene in Ectropis oblique

DU Qin, XIN Zhaojun, YANG Ziwei, LI Xiwang, CHEN Zongmao, SUN Xiaoling   

  1. Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture/Institute of Tea Research, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China
  • Received:2014-05-08 Revised:1900-01-01 Online:2015-02-08 Published:2015-02-08

摘要: 昆虫脂肪酶可参与脂类消化、吸收和运输,并能介导植物的抗病虫功能。本研究通过茶尺蠖唾液腺转录组数据库发掘到该基因片段,采用RACE技术克隆获得了一条茶尺蠖脂肪酶合成基因序列,命名为EoL2,并检测了其在茶尺蠖不同生育期及不同组织部位中的相对表达水平。研究结果表明,EoL2的cDNA序列全长1310 bp,开放阅读框为1041 bp,编码一个包含346个氨基酸的蛋白,推测该蛋白相对分子量为37.96 kD,推测等电点(PI)为8.41,具有脂肪酶典型的GXSXG丝氨酸活性保守序列,其氨基酸序列与家蚕相似度最高,为57%。实时荧光定量PCR结果表明,EoL2在卵中的表达量显著低于其它虫态;在1龄幼虫期的相对表达水平显著高于2~5龄;在成虫中的表达量显著高于蛹;在血淋巴中的表达量显著高于中肠、表皮、脂肪体和唾液腺。

Abstract: Herbivore lipases participate in lipid digestion, absorption and transportation; and mediate interactions between plant and pathogens or herbivore insects as well. Based on transcriptome analysis of Ectropis oblique salivary glands, a putative gene encoding a lipase gene was obtained by rapid amplification of cDNA ends for the first time, which was named as EoL2. The relative expression levels of EoL2 in different stages and tissues were measured. The results show that EoL2 contains an open reading frame of 1041 bp which encodes a protein of 346 amino acid with a putative molecular mass of 37.96 kD and an isoelectric point of 8.41. The deduced amino acid sequence contains a serine conserved sequence GXSXG. According to the alignment, EoL2 shares 57% similarity of amino acid sequence with Bombyx mori L1. The results of real-time quantitative PCR show that EoL2 expression was lower in eggs than in other stages, higher in the first instar than in other instars, higher in adults than in pupae, and higher in hemolymph than in midgut, cuticula, fat body and salivary gland.

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