拟南芥抗病相关基因<em>T1N6_22</em>互作蛋白的酵母双杂交鉴定

doi:10.7668/hbnxb.2018.02.001

华北农学报 ›› 2018, Vol. 33 ›› Issue (2): 1-7. doi: 10.7668/hbnxb.2018.02.001

所属专题： 生物技术；

拟南芥抗病相关基因T1N6_22互作蛋白的酵母双杂交鉴定

赵亚婷¹, 邢红侠¹, 庞茜¹, 郑旭¹, 张靖¹, 瓮巧云^1,2, 邢继红¹, 董金皋¹

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

收稿日期:2017-12-26 出版日期:2018-04-28
通讯作者: 邢继红(1977-),女,河北东光人,教授,博士,主要从事分子植物病理学研究;董金皋(1963-),男,河北平乡人,教授,博士,主要从事植物病理学研究。
作者简介:赵亚婷(1993-),女,河北邯郸人,硕士,主要从事植物抗性生理学研究。
基金资助:
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）

Identification of Interacting Proteins of Resistance-related Gene T1N6_22 in Arabidopsis thaliana by Yeast Two-hybrid

ZHAO Yating¹, XING Hongxia¹, PANG Xi¹, ZHENG Xu¹, ZHANG Jing¹, WENG Qiaoyun^1,2, XING Jihong¹, DONG Jingao¹

1. Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, Mycotoxin and Molecular Plant Pathology Laboratory of Hebei Agricultural University, Baoding 071001, China;
2. College of Agriculture and Forestry Science and Technology, Hebei North University, Zhangjiakou 075000, China

Received:2017-12-26 Published:2018-04-28

摘要/Abstract

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。

关键词: 拟南芥, T1N6_22, 酵母双杂交, 互作蛋白

Abstract: 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.

Key words: Arabidopsis thaliana, T1N6_22, Yeast two-hybrid, Interacting protein

中图分类号:

Q945.8
Q78

赵亚婷, 邢红侠, 庞茜, 郑旭, 张靖, 瓮巧云, 邢继红, 董金皋. 拟南芥抗病相关基因T1N6_22互作蛋白的酵母双杂交鉴定[J]. 华北农学报, 2018, 33(2): 1-7. doi: 10.7668/hbnxb.2018.02.001.

ZHAO Yating, XING Hongxia, PANG Xi, ZHENG Xu, ZHANG Jing, WENG Qiaoyun, XING Jihong, DONG Jingao. Identification of Interacting Proteins of Resistance-related Gene T1N6_22 in Arabidopsis thaliana by Yeast Two-hybrid[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(2): 1-7. doi: 10.7668/hbnxb.2018.02.001.

http://www.hbnxb.net/CN/Y2018/V33/I2/1

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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

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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

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