松墨天牛谷氨酰胺6-磷酸果糖酰胺转移酶基因克隆与表达  

Isolation and Expression of Glutamine-Fructose-6-Phosphate Aminotransferase Gene from Monochamus alternatus

作　　者：蔡紫玲 吴华俊[1] 林同[1] 

机构地区：[1]华南农业大学林学与风景园林学院,广东广州510642

出　　处：《西南农业学报》2016年第9期2131-2137,共7页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金(31470653);广东省自然科学基金(1414050001666)

摘　　要：谷氨酰胺6-磷酸果糖酰胺转移酶(glutamine-fructose-6-phosphate aminotransferase,GFAT)是己糖胺通路(HBP)的限速酶,参与几丁质的合成和蛋白质的糖基化。本研究利用RACE克隆了松墨天牛GFAT基因(Ma GFAT),Gen Bank登录号为KT362367,其长度为2624 bp,开放阅读框为2061 bp,编码686个氨基酸。序列对比分析显示Ma GFAT与赤拟谷盗相似性最高,为88%,在系统发育树上位于同一分支。采用RT-q PCR方法检测Ma GFAT在松墨天牛不同组织及不同发育时期的表达情况,结果表明:Ma GFAT在各虫态中,成虫中的表达量最高;在成虫部位中,翅的表达量最高;在幼虫各组织中,体壁的表达量最高。本文为研究GFAT结构与功能的关糸奠定基础,为阐明GFAT在松墨天牛几丁质合成中的作用提供参考。Glutamine-fructose-6-phosphate aminotransferase（ GFAT） was a rate-limiting enzyme of hexosamine biosynthetic pathway,which had a regulatory role in chitin synthase and protein glycosylation. The GFAT gene of Monochamus alternatus（ Ma GFAT,Gen Ban K accession number is KT362367） by RACE was cloned and sequenced,and its length was 2624 bp,including 2061 bp open reading frame encoding686 amino acids. The sequence analysis showed that Ma GFAT had a high similarity with Tribolium castaneum at 88 % amino acid level,and both of the GFAT of M. alternatus and T. castaneum were in the same branch of the phylogenetic tree. The Ma GFAT expression in different tissues and development period by RT-q PCR were detected. The results showed that Ma GFAT had the highest expression in adults among developmental periods,in wings among body parts of adults and in body wall among all larvae tissues. This result would provide references for investigating the relation between structure and function of GFAT and clarifying the potential role of GFAT which played in biosynthesis of chitin in M. alternatus.

关 键 词：松墨天牛 谷氨酰胺6-磷酸果糖酰胺转移酶 基因克隆 RT-q PCR 

分 类 号：Q966[生物学—昆虫学]