《植物生理学报》 2015, 51(11): 1895-1900

苹果转录因子MdHB1自激活作用及其互作蛋白分析

刘艳利¹, 姜永华¹, 王豪杰¹, 戚英伟², 刘翠华¹, 任小林^1,*

¹西北农林科技大学园艺学院, 陕西杨凌712100; ²宁夏大学农学院, 银川750021

通信作者：任小林；E-mail: renxl@nwsuaf.edu.cn；Tel: 029-87099217

摘　要：

筛选苹果转录因子MdHB1的互作蛋白, 可以为研究苹果生长发育过程中各种信号分子网络提供线索和依据。本文以苹果品种‘皇家嘎啦’为试材, 应用SMART技术构建酵母双杂交cDNA文库。分析MdHB1的转录活性区域并验证其对Y2HGold的细胞毒性作用, 利用诱饵载体pGBKT7-MdHB1-N进行文库筛选, 获得了24个阳性克隆, 测序分析后得到9个无重复的MdHB1互作蛋白, 其涉及光合作用、病虫害防御、逆境胁迫等过程。

关键词：互作蛋白; 酵母双杂交文库; SMART技术; MdHB1

收稿：2015-06-29   修定：2015-10-27

资助：农业部现代农业技术体系苹果专项资金(MATS)。

Self Activation and Interaction Protein Analysis of MdHB1 Transcription Factor in Apple

LIU Yan-Li¹, JIANG Yong-Hua¹, WANG Hao-Jie¹, QI Ying-Wei², LIU Cui-Hua¹, REN Xiao-Lin^1,*

¹College of Horticulture, Northwest A & F University, Yangling, Shaanxi 712100, China; ²Agricultural College, Ningxia University, Yinchuan 750021, China

Corresponding author: REN Xiao-Lin; E-mail: renxl@nwsuaf.edu.cn; Tel: 029-87099217

Abstract:

The screening of protein interactions with our target protein MdHB1 in apples provides clues and evidences to the study of molecular network in growth and development. This experiment used ‘Royal Gala’ as the testing material and SMART technology was used to construct the yeast two-hybrid cDNA library. We analyzed that MdHB1 protein had obvious self-activation activity, while the N-terminus of MdHB1 (excluding with transcription activity of CS2 structure domain) had no self-activation activity and toxicity. The quality of the library was tested and used for subsequent screening. Finally, we screened 9 positive dots using pGBKT7-MdHB1-N. Function annotation showed that these candidate interacting proteins were related to photosynthesis, defense reaction and resistance.

Key words: protein interactions; yeast two-hybrid library; SMART technology; MdHB1