八字地老虎和粘虫蜕皮激素接受子3(HR3)基因cDNA序列的克隆与序列分析

Molecular cloning and sequences analysis of molt hormone receptor 3 ( HR3 ) cDNA from Agrotis c-nigrum and Mythimna separata

蜕皮激素接受子3(hormone receptor 3,HR3),是一种蜕皮调节转录因子,调控蜕皮过程中相关基因的表达,是蜕皮级联反应中的关键因子。本文以八字地老虎Agrotisc-nigrumL.和粘虫Mythimna separata Walker预蛹期幼虫为材料,分别提取总RNA,利用RT-PCR和cDNA末端快速扩增技术(RACE),分别扩增得到2种昆虫蜕皮激素接受子3(HR3)的5′非编码区和完整开放读码框在内的cDNA序列,其中八字地老虎的HR3 cDNA序列含有1729个碱基,包括一个1533个碱基的开放阅读框,编码一个含510个氨基酸的蛋白,分子量约为57.5ku。粘虫的HR3cDNA序列含有1743个碱基,包括一个1536个碱基的开放阅读框,编码一个含511个氨基酸的蛋白,分子量约为57.9ku。这2种昆虫HR3 cDNA序列推导的氨基酸序列均具有昆虫核受体超家族特征性结构域,与其他昆虫,尤其是鳞翅目昆虫的蜕皮激素接受子3的氨基酸序列高度同源。获得的基因cDNA序列已经登录GenBank并获得登录号,八字地老虎HR3登录号为GU188853,粘虫HR3登录号为GU188854。

Insect molt hormone receptor 3 （ HR3 ） is a molt-regulating transcription factor that plays an important role in regulating insect molt. Total RNA was isolated from the prepupae of Agrotis c-nigrum L. and Mythimna separata Walker. The cDNA sequences were cloned by RT-PCR and rapid amplification of cDNA ends （RACE）. The cDNA sequence of A. c-nigrum HR3 was 1 729 base pairs in length and contained an open reading frame of 1 533 base pairs coding for a polypeptide of 510 amino acid residues with a predicted molecular weight of 57. 5 ku. The cDNA sequence of M. separate HR3 was 1 743 base pairs in length and contained an open reading frame of 1 536 base pairs coding for a polypeptide of 511 amino acid residues with a predicted molecular weight of 57. 9 ku. The predicted protein of both insects had the characteristics of the nuclear receptor superfamily and shared extensive similarities with those from other insects,especially the Lepidoptera. The cDNA sequences have been deposited in GenBank with accession numbers GU188853 for A. c-nigrum HR3 and GU188854 for M. separata HR3.