龙眼成花逆转花芽α-tubulin基因的克隆与原核表达  

Cloning and Prokaryotic Expression of α-tubulinGene in Longan Floral Reversion Buds

作　　者：游向荣[1,2,3] 黄榕辉[2] 王喜军[2] 黄春梅[2] 梁文裕[2] 陈伟[1,2] 

机构地区：[1]福建农林大学作物遗传育种与综合利用教育部重点实验室,福州350002 [2]福建农林大学生命科学学院,福州350002 [3]广西农业科学院农产品加工研究所,南宁530007

出　　处：《热带亚热带植物学报》2011年第1期63-68,共6页Journal of Tropical and Subtropical Botany

基　　金：国家自然科学基金项目(30571293);福建省自然科学基金项目(2007J0045);教育部博士点基金(200803890009)资助

摘　　要：运用蛋白质组学方法比较龙眼(Dimocarpus longanLour.)正常成花和成花逆转花芽的差异蛋白质组,并应用RACE方法克隆其中上调表达的α-微管蛋白基因α-tubulin,获得一段长度为1641 bp的cDNA,其中包括1个1350 bp的开放阅读框[GenBank登录号:FJ479617(GI:218202929)]。将α-tubulin全长cDNA在大肠杆菌中表达,获得1个约49.6 kD的外源蛋白,经Western blotting验证为α-微管蛋白。RT-PCR和Western blotting分别检测了α-tubulin在转录和翻译水平上的表达,结果表明,α-微管蛋白在成花逆转的龙眼花芽中上调表达,可能是逆转花芽形态差异表现的原因之一。The differential proteins of floral reversion buds at different stages in longan（Dimocarpus longan Lour.） were compared using proteomics method.The results showed that α-tubulin could up-regulate in floral reversion buds of longan,and its gene α-tubulin was cloned using RACE method.A full length of 1641 bp cDNA,with a 1350 bp open reading frame,was obtained（GenBank accession number： FJ479617）.When α-tubulin gene was transformed into E.coli and expressed,a 49.6 kD heterologous protein verified as α-tubulin by Western blotting was obtained.Different expression of α-tubulin at transcription and translation levels using RT-PCR and Western blotting,respectively,showed that α-tubulin up-regulated in longan floral reversion buds.It was one of reasons that floral reversion buds were different from normal flowering buds.

关 键 词：龙眼 花芽 α-tubulin基因 克隆 原核表达 

分 类 号：S667.2[农业科学—果树学]