Position: Home > Articles > Screening PbTMT4 interaction factor by yeast two-hybrid system in pear
Hubei Agricultural Sciences
2019
(24)
240-245
采用酵母双杂交系统筛选梨PbTMT4互作因子
作 者：
程寅胜;杨立;陈健秋;张绍铃;伍涛;张虎平
单 位：
湖北省农业科学院果树茶叶研究所;南京农业大学园艺学院
关键词：
PbTMT4;酵母双杂交系统;cDNA文库;互作因子
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。
译 名：
Screening PbTMT4 interaction factor by yeast two-hybrid system in pear
作 者：
CHENG Yin-sheng;YANG Li;CHEN Jian-qiu;ZHANG Shao-ling;WU Tao;ZHANG Hu-ping;Institute of Fruit Tree Tea,Hubei Academy of Agricultural Sciences;College of Horticulture,Nanjing Agricultural University;
关键词：
PbTMT4;;yeast two-hybrid system;;cDNA library;;interaction factor
摘 要：
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 system, the constructed recombinant plasmid pDHB1-PbTMT4 was used as a bait plasmid to transform yeast strain NMY51 by PEG/Li Ac 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 nontoxic 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.
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