采用酵母双杂交系统筛选梨PbTMT4互作因子

doi:10.14088/j.cnki.issn0439-8114.2019.24.058

湖北农业科学 ›› 2019, Vol. 58 ›› Issue (24): 240-245.doi: 10.14088/j.cnki.issn0439-8114.2019.24.058

采用酵母双杂交系统筛选梨PbTMT4互作因子

程寅胜¹, 杨立¹, 陈健秋², 张绍铃², 伍涛¹, 张虎平²

1.湖北省农业科学院果树茶叶研究所,武汉 430064;
2.南京农业大学园艺学院,南京 210095

收稿日期:2019-09-16 出版日期:2019-12-25 发布日期:2019-12-25
通讯作者: 张虎平,副教授,主要从事果树栽培与育种研究,（电子信箱）hpzhang@njau.edu.cn; 伍涛,副研究员,主要从事梨树栽培生理研究,（电子信箱）wutaoga556@163.com。
作者简介:程寅胜（1991-）,男,安徽桐城人,研究实习员,硕士,主要从事果树分子生物学和果实品质研究,（电话）15295595528（电子信箱） chengyinsheng1991@163.com
基金资助:
中央高校基本科研业务费专项（KYZ201834）; 湖北省自然科学基金项目（2018CFB285）

Screening PbTMT4 interaction factor by yeast two-hybrid system in pear

CHENG Yin-sheng¹, YANG Li¹, CHEN Jian-qiu², ZHANG Shao-ling², WU Tao¹, ZHANG Hu-ping²

1.Institute of Fruit Tree Tea,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;
2.College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China

Received:2019-09-16 Online:2019-12-25 Published:2019-12-25

摘要/Abstract

摘要： 为筛选梨糖转运相关蛋白PbTMT4互作因子,基于已经建立好的砀山酥梨果实发育时期的cDNA文库,利用Matchmaker^TM Gold酵母双杂交系统,以构建的重组质粒pDHB1-PbTMT4为诱饵质粒,采用PEG/LiAc法转化酵母菌株NMY51。利用表型筛选法检测PbTMT4的自激活作用和毒害作用,利用氨基酸缺陷型培养基筛选PbTMT4互作因子。结果表明,诱饵蛋白PbTMT4在酵母双杂交系统中无毒性和自激活作用。通过酵母双杂交筛选以及测序和序列比对分析,共获得13个与PbTMT4蛋白相互作用的宿主蛋白。

关键词: PbTMT4, 酵母双杂交系统, cDNA文库, 互作因子

Abstract: To screen for the interaction factor of PbTMT4, a pear sugar transport related gene, based on the established cDNA library of Dangshan Suli pear fruit development period, using Matchmaker^TM Gold yeast two-hybrid syste m, the constructed recombinant plasmid pDHB1-PbTMT4 was used as a bait plasmid to transform yeast strain NMY51 by PEG/LiAc method. The phenotypic screening method was used to detect the self-activation and toxic effects of PbTMT4. The PbTMT4 gene interaction factor was screened using an amino acid-deficient medium. The results showed that the bait protein PbTMT4 was non-toxic and self-activating in the yeast two-hybrid system. Through yeast two-hybrid screening and sequencing and sequence alignment analysis, a total of 13 host proteins interacting with PbTMT4 protein were obtained.

Key words: PbTMT4, yeast two-hybrid system, cDNA library, interaction factor

中图分类号:

S661.2

程寅胜, 杨立, 陈健秋, 张绍铃, 伍涛, 张虎平. 采用酵母双杂交系统筛选梨PbTMT4互作因子[J]. 湖北农业科学, 2019, 58(24): 240-245.

CHENG Yin-sheng, YANG Li, CHEN Jian-qiu, ZHANG Shao-ling, WU Tao, ZHANG Hu-ping. Screening PbTMT4 interaction factor by yeast two-hybrid system in pear[J]. HUBEI AGRICULTURAL SCIENCES, 2019, 58(24): 240-245.

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采用酵母双杂交系统筛选梨PbTMT4互作因子

Screening PbTMT4 interaction factor by yeast two-hybrid system in pear

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