VDAC3与ABA受体蛋白相互作用的验证以及初步研究  

Authentication and preliminary study of interactions between VDAC3 and RCAR1,RCAR3

作　　者：蔡黎[1] 李德款[1] 刘志斌[1] 冯俊[1] 陈存[1] 李旭锋[1] 杨毅[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2013年第5期1097-1103,共7页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(30971557;31171586)

摘　　要：酵母双杂交技术在拟南芥cDNA文库中筛选得到了与ABA受体蛋白RCAR1,RCAR3均有相互作用的蛋白质VDAC3,并已获得VDAC3基因的全长.VDAC3蛋白包含有3个结构域,分别为Porin3结构域,DUF3442结构域和SEG片段,现根据结构域分布,将VDAC3截短为包含DUF3442的VDAC3N以及包含SEG的VDAC3C两段.将VDAC3全长以及分别截短的169个氨基酸(VDAC3N)及105个氨基酸(VDAC3C)的片段进行酵母转化实验,与RCAR1或RCAR3共同转化后,VDAC3,VDAC3N的共转酵母能够在缺陷型培养基上生长,并且使X-Gal显色为蓝色,而VDAC3C则不能.这验证了全长的VDAC3与RCAR1,RCAR3的存在相互作用,并发现决定VDAC3蛋白与RCAR1,RCAR3是相互作用的结构域位于N端的DUF3442结构域.VDAC3 was screened out as interacting protein with abscisic acid（ABA） receptors RCAR1 and RCAR3 in Arabidopsis thialiana cDNA library by the yeast two hybrid system before. VDAC3 contains three domains. Porin3 domain,DUF3442 domain and SEG fragment. In this research VDAC3 was cut into two part, VDAC3 N which contains DUF3442 domain while VDAC3 C which contains SEG frag ment. Cotransform BD-RCAR1, BD-RCAR3 and full length AD-VDAC3, AD-VDAC3 N （507bp） into yeasts. The reconstructedyeasts can grow on themediumSD/-His/-Trp/-Leu/-Ade and turn blue in the presence of X-Gal while yeasts cotransformed BD-RCAR1, BD-RCAR3and AD-VDAC3 C（318 bp）can＇t. These experiments proved thatthe full length VDAC3 indeed interacts with RCAR1 and RCAR3,and al so N-termini DUF3442 domainpossesses the interactions.

关 键 词：ABA受体蛋白 VDAC3 酵母双杂交系统 

分 类 号：Q945[生物学—植物学]