拟南芥抗病相关基因T1N6_22互作蛋白的酵母双杂交鉴定  

Identification of Interacting Proteins of Resistance-related Gene T1N6-22 in Arabidopsis thaliana by Yeast Two-hybrid

作　　者：赵亚婷[1] 邢红侠 庞茜[1] 郑旭[1] 张靖[1] 瓮巧云[1,2] 邢继红[1] 董金皋[1] ZHAO Yating;XING Hongxia;PANG Xi;ZHENG Xu;ZHANG Jing;WENG Qiaoyun;XING Jihong;DONG Jingao(Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology,Mycotoxin and Molecular Plant Pathology Laboratory of Hebei Agricultural University,Baoding 071001,China;College of Agriculture and Forestry Science and Technology,Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北省植物生理与分子病理学重点实验室,河北农业大学真菌毒素与植物分子病理学实验室,河北保定071001 [2]河北北方学院农林科技学院,河北张家口075000

出　　处：《华北农学报》2018年第2期1-7,共7页Acta Agriculturae Boreali-Sinica

基　　金：河北省自然科学基金项目(C2014405010);2017年大学生创新创业训练计划项目(201710086012)。

摘　　要：利用酵母双杂交技术,鉴定拟南芥抗病相关基因T1N6-22的互作蛋白,为进一步明确T1N6-22基因调控拟南芥抗病的分子机制奠定基础。利用Gateway技术,构建T1N6-22基因及其候选互作蛋白基因的酵母双杂交载体AD-T1N6-22、AD-AT1G06050、AD-AT1G21400、AD-AT2G19480、BD-T1N6-22、BD-AT1G06050、BD-AT1G21400和BD-AT2G19480。将空载的AD载体分别与BD-T1N6-22、BD-AT1G06050、BD-AT1G21400和BD-AT2G19480载体组合共同转化酵母感受态细胞,检测各基因的自激活活性,发现T1N6-22、AT1G21400、AT2G19480基因无自激活活性,AT1G06050基因有自激活活性。将AD-T1N6-22载体分别与BD-AT1G06050、BD-AT1G21400和BD-AT2G19480载体组合,BD-T1N6-22载体分别与AD-AT1G06050、AD-AT1G21400、AD-AT2G19480载体组合,进行酵母双杂交试验。结果发现,AD-T1N6-22与BD-AT1G06050、BD-AT1G21400和BD-AT2G19480,BD-T1N6-22与AD-AT1G06050、AD-AT1G21400、AD-AT2G19480共转化的酵母细胞在二缺(-Leu/-Trp)、三缺(-Leu/-Trp/-His)和四缺(-Leu/-Trp/-His/-Ade)培养基上均可以生长,表明T1N6-22与AT1G06050、AT1G21400和AT2G19480在酵母细胞中直接互作。The objective of this study is to identify interacting proteins of resistance-related gene T1N6_22 in Arabidopsis thaliana,and to provide a basis for clarifying the regulation mechanism of the T1N6_22 gene in Arabidopsis resistance.The yeast two-hybrid vectors of the T1N6_22 gene and its candidate interacting protein gene,including AD-T1N6_22,AD-AT1G06050,AD-AT1G21400,AD-AT2G19480,BD-T1N6_22,BD-AT1G06050,BD-AT1G21400,and BD-AT2G19480 were constructed by Gateway method.In order to detect self-activation of T1N6_22,AT1G06050,AT1G21400,and AT2G19480 genes,the AD vector was combined with BD-T1N6_22,BD-AT1G06050,BD-AT1G21400,and BD-AT2G19480 respectively,and transformed into yeast competent cells.The results showed that AT1G06050 had self-activation activity and T1N6_22,AT1G21400,AT2G19480 didn′t have self-activation activity.For yeast two hybrid,the AD-T1N6_22 vector was combined with BD-AT1G06050,BD-AT1G21400,BD-AT2G19480 vectors,respectively,and the BD-T1N6_22 vector was combined with AD-AT1G06050,AD-AT1G21400,AD-AT2G19480 vectors,respectively.It was found that yeast colonies which were co-transformed with AD-T1N6_22 and BD-AT1G06050,BD-AT1G21400 or BD-AT2G19480,BD-T1N6_22 and AD-AT1G06050,AD-AT1G21400 or AD-AT2G19480 grew well in both-Leu/-Trp and-Leu/-Trp/-His and-Leu/-Trp/-His/-Ade media.Yeast two-hybrid results indicated that the T1N6_22 gene directly interacted with AT1G06050,AT1G21400 and AT2G19480 in yeast.

关 键 词：拟南芥 T1N6_22 酵母双杂交 互作蛋白 

分 类 号：Q945.8[农业科学—植物病理学] Q78[生物学—植物学]