[1]周汐,张璟曜,姚敏磊,等.大豆酵母杂交cDNA文库的构建及GmSPX3互作蛋白的筛选[J].大豆科学,2016,35(01):18-24.[doi:10.11861/j.issn.1000-9841.2016.01.0018]
　ZHOU Xi,ZHANG Jing-yao,YAO Min-lei,et al.Construction of Yeast Hybrid cDNA Library and Screening of Interaction Proteins of GmSPX3 in Soybean[J].Soybean Science,2016,35(01):18-24.[doi:10.11861/j.issn.1000-9841.2016.01.0018]
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大豆酵母杂交cDNA文库的构建及GmSPX3互作蛋白的筛选

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年01期 页码: 18-24 栏目: 出版日期: 2016-01-20
Title:
Construction of Yeast Hybrid cDNA Library and Screening of Interaction Proteins of GmSPX3 in Soybean
作者:
周汐; 张璟曜; 姚敏磊; 谢凤斌; 盖钧镒; 杨守萍
南京农业大学 大豆研究所/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室/作物遗传与种质创新国家重点实验室，江苏 南京 210095
Author(s):
ZHOU Xi;  ZHANG Jing-yao;  YAO Min-lei;  XIE Feng-bin;  GAI Jun-yi;  YANG Shou-ping
Soybean Research Institute/National Center for Soybean Improvement/MOA Key Laboratory for Biology and Genetic Improvement of Soybean(General)/National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
关键词:
大豆; GmSPX3基因; Gateway技术; 酵母杂交cDNA文库; 酵母双杂交
Keywords:
Soybean［Glycine max (L) Merr］;  GmSPX3 gene;  Gateway technology;  Yeast hybrid cDNA library;  Yeast two-hybrid
DOI:
10.11861/j.issn.1000-9841.2016.01.0018
文献标志码:
A
摘要:
为研究GmSPX3在大豆低磷胁迫响应中的作用机制，寻找与GmSPX3发生相互作用的蛋白质，利用Gateway技术构建了低磷大豆根系酵母杂交cDNA文库，并使用pGBKT7-GmSPX3融合蛋白为诱饵，采用酵母双杂交方法筛选酵母杂交cDNA文库，经过显色反应验证后获得21个阳性克隆，通过生物信息学分析，选择2个转录因子编码基因GmUNE12和GmPosF21作为候选基因进行克隆，并分别构建AD载体pGADT7-GmUNE12 和 pGADT7-GmPosF21，与诱饵pGBKT7-GmSPX3进行一对一互作验证，结果证明GmUNE12和GmPosF21蛋白均与GmSPX3诱饵蛋白发生了相互作用。
Abstract:
To study the mechanism of GmSPX3 in response to low phosphorus stress and search for the interaction proteins of GmSPX3 in soybean, the present research constructed yeast hybrid cDNA library from the soybean root treated with low phosphorus by gateway technology Furthermore, yeast hybrid cDNA library was screened through the method of yeast two-hybrid using pGBKT7-GmSPX3 fusion protein as bait. After color reaction of validation, 21 positive clones were obtained. Two transcription factors GmUNE12 and GmPosF21 were chosen as the candidate genes for cloning by bioinformatics analysis. Two AD vectors of pGADT7-GmUNE12 and pGADT7-GmPosF21 were constructed. The results of one to one interaction demonstrated that GmUNE12 and GmPosF21 proteins interacted with GmSPX3 bait protein, respectively

参考文献/References:

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备注/Memo

基金项目：国家转基因重大专项（2011ZX08004-005，2013ZX08004-005，2014ZX08004-005）；国家高技术研究发展计划“863计划”（2011AA10A105）；国家重点基础研究发展计划“973计划”（2011CB109301）；教育部长江学者和创新团队发展计划（PCSIRT13073）；江苏省现代作物生产协同创新中心（JCIC-MCP）。

第一作者简介：周汐（1989-），女，硕士，主要从事大豆遗传育种研究。E-mail：ciel891259@gmail.com。
通讯作者：杨守萍（1967-），女，博士，教授，主要从事大豆遗传育种研究。E-mail：spyung@126.com。

更新日期/Last Update: 2016-01-28