文档介绍：
粘虫毒蕈碱型乙酰胆碱受体基因的克隆与
发育表达分析#
胡耿,姜鸣,吕淑敏**
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(西北农林科技大学植保资源与害虫治理教育部重点实验室,陕西杨凌,712100)
摘要:本文应用 RT-PCR 结合 RACE 技术获得了粘虫 Mythimna separata 毒蕈碱型乙酰胆碱
受体(muscarine acetylcholine receptor, mAchR)基因的全长 cDNA 序列 2369 bp,开放阅读
框 1806 bp,5’端非编码区 274 bp,3’端非编码区 290 bp,编码 602 个氨基酸残基,分子
量为 kDa,理论等电点 pI=。同源性分析表明该蛋白与意大利蜜蜂 Apis mellifera
mAchR 的氨基酸序列最大一致性达 56%,与双翅目昆虫 mAchR 的亲缘关系最近。采用
SYBR Green I 实时定量 RT-PCR 方法,检测粘虫各发育阶段 mAchR 基因 mRNA 的相对表
达量,结果显示,mAchR 基因在粘虫整个发育过程中均有表达,以卵期及成虫期表达量最
高,幼虫期随着龄期的增长表达量逐渐降低,蛹期有所升高,成虫期雌雄虫表达差异极显
著,雌虫的表达量可达雄虫的 20 倍以上。以上结果说明粘虫 mAchR 可能在卵期发育中起
到重要作用,雌成虫期的高表达可能与其交配或产卵行为相关。
关键词:粘虫,毒蕈碱型乙酰胆碱受体,生物信息学,发育,实时定量 RT-PCR
中图分类号:Q965
Cloning and developmental expression analysis of a
muscarine acetylcholine receptor gene from Mythimna
separate (Lepidoptera: Noctuidae)
HU Geng, JIANG Ming, LV Shumin
(Key Laboratory of Plant Protection Resources
and Pest Management,Ministry of Education,Northwest A&F University,Yangling,Shaanxi
712100,China)
Abstract: The full-length cDNA of muscarine acetylcholine receptor (mAchR) was cloned from
Mythimna separata by RT-PCR & RACE technology. It is 2369 bp with an ORF of 1806 bp,
containing a 5'-untraslated region (5'-UTR) of 274 bp and a 3'-UTR of 290 bp. It encodes a deduced
protein of 602-amino acids residues, with a predicted molecular mass of kDa and isoeletric point
(pI) of . The putative amino acid shares 56% similarity with Apis mellifera mAchR protein, and
ic analysis showed that it is mostly related to that of Diptera insects. Real-time PCR results
indicated that M. separata mAchR was expressed in all development stages of M. separate, with the
highest expression level in egg and adult stages. It is gradually reduced from the 1st instar to the 6th
instar, with a little increase in the pupae stage. In adult stage, there is significant difference between the
male and female adult, the expression level of female is 20 times more than that of male. These results
r