中国农业科技导报 ›› 2016, Vol. 18 ›› Issue (6): 65-71.DOI: 10.13304/j.nykjdb.2016.233

• 生物技术 生命科学 • 上一篇    下一篇

东亚飞蝗储存蛋白基因LmiHex-2的克隆与表达分析

李艳丽,郝金凤,赵学乾,印红*#br#   

  1. 河北大学生命科学学院, 河北省动物系统学与应用重点实验室, 河北 保定 071002
  • 收稿日期:2016-04-21 出版日期:2016-12-15 发布日期:2016-07-04
  • 通讯作者: 印红,教授,博士,主要从事昆虫分子系统发育研究。E-mail:yinhong@hbu.edu.cn
  • 作者简介:李艳丽,硕士研究生,主要从事昆虫分子系统发育研究。 E-mail: 1305561669@qq.com。
  • 基金资助:
    国家自然科学基金项目(31272293)资助。

Cloning and Expression Analysis of a Hexamerin Gene LmiHex-2 from Locusta migratoria manilensis (Acridoidea: Oedipodidae)

LI Yan-li, HAO Jin-feng, ZHAO Xue-qian, YIN Hong*   

  1. Key Laboratory of Zoological Systematics and Application, College of Life Sciences, Hebei University, Hebei Baoding 071002, China
  • Received:2016-04-21 Online:2016-12-15 Published:2016-07-04

摘要: 昆虫储存蛋白是在昆虫体内普遍存在的一种特异性淋巴蛋白,在昆虫的生长发育过程中发挥着重要作用。为分析东亚飞蝗Locusta migratoria manilensis储存蛋白基因LmiHex-2的结构和时空表达特性,通过RACE技术获得了LmiHex-2基因2 171 bp全长cDNA序列,其中开放阅读框2 022 bp,编码673个氨基酸,预测分子量78.79 kDa。序列比对结果显示LmiHex-2基因cDNA序列与直翅目蝗总科储存蛋白基因cDNA序列的相似性达到82%~89%。RT-PCR结果显示LmiHex-2基因在东亚飞蝗卵和若虫阶段均有表达,5龄6 d时表达量最大;成虫中脂肪体表达量最高。通过贝叶斯法构建的系统发育树显示直翅目储存蛋白为单系群,昆虫血蓝蛋白与昆虫储存蛋白聚为姐妹群,二者均由甲壳纲血蓝蛋白进化而来。

关键词: 东亚飞蝗, 储存蛋白, 基因表达, 序列分析, 进化树

Abstract: Hexamerins are large hemolymph-proteins that accumulate in insects during the late larval stages and play important roles in insect development and growth. To investigate the structure and expression profiles of Locusta migratoria manilensis hexamerin (LmiHex-2), the complete cDNA (2 171 bp) of LmiHex-2 was obtained by RACE method. The LmiHex-2 contained an open reading frame of 2 022 bp, which encoded 673 amino acids with a predicted molecular mass of 78.79 kDa. Sequence alignment revealed that the LmiHex-2 shared an identity of 82%~89% with hexamerins from other Acridoidea species. RT-PCR results showed that LmiHex-2 gene was expressed both in nymphs and adults. After 6 d 5th instar nymphs, the relative expression quantity of LmiHex-2 reached a maximum. In adult males and females, the fat body exhibited the highest level of LmiHex-2 transcripts. The phylogenetic tree showed that Orthoptera hexamerins was a monophyletic group, the Insecta hexamerins and the  Insecta  hemocyanins, which evolved from Crustacea hemocyanins and form a sister group.

Key words: Locusta migratoria manilensis, hexamerin, gene expression, sequence analysis, phylogenetic tree