利用酵母双杂交系统筛选花生AhMYB44互作蛋白
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1江苏省农业科学院经济作物研究所,南京 210014,2 农业农村部油料作物生物学与遗传育种重点实验室,武汉 430000

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国家现代农业产业技术体系建设项目(CARS-13);江苏省农业科技自主创新资金项目(cx(18)2015);农业农村部油料作物生物学与遗传育种重点实验室开放课题项目(KF2019011)


Yeast Two-Hybrid Screening of the Proteins Interacting with AhMYB44 from Peanut
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1 Jiangsu Academy of Agricultural Sciences,Nanjing 210014;2 Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430000

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Special Fund for Modern Agroindustry Technology Research System of China (CARS-13), Independent Innovation Fund of Agricultural Science and Technology of Jiangsu Province(cx(18)2015), Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, P.R. China(KF2019011)

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    摘要:

    前期根据花生干旱转录组测序结果克隆了一个胁迫诱导表达基因 AhMYB44 的全长序列。为进一步研究基因的功能,本 研究利用酵母双杂交系统进行其互作蛋白的筛选。结果显示:根据基因序列构建的诱饵载体 pGBKT7-AhMYB44 本身无毒性,且 无自激活活性;经与花生干旱及高盐胁迫相关的 cDNA 文库杂交、筛选、鉴定出 16 个插入序列,它们对应的同源蛋白功能涉及植物 转录调控、生长发育、信号转导、生物及非生物胁迫响应等多个方面;其中 46 号候选互作蛋白与非生物胁迫密切相关,结构域分析 属于钙依赖性磷脂结合蛋白(膜联蛋白,ANNs),植物膜联蛋白已被证实参与多种胁迫响应过程。因此推测 AhMYB44 可能通过胞 内钙信号调控花生胁迫响应过程。这也为今后深入开展 AhMYB44 的功能及作用机制研究提供了参考依据。

    Abstract:

    We previously cloned the full-length coding sequence of the gene AhMYB44,which showed a stress-induced expression from peanut drought transcriptome sequencing datasets. In order to investigate the function of AhMYB44,a yeast two-hybrid system was deployed for identifying its interacting proteins. The results showed that the bait vector pGBKT7-AhMYB44 was not toxic and showed no self-activation activity. Sixteen coding sequences were identified by screening of cDNA libraries which were constructed from a drought and high-salt stress of peanut. We further conducted an in-silico analysis of their homologous proteins,such as plant transcriptional regulation,growth and development,signal transduction,biotic and abiotic stress responses. Out of that,the candidate gene(No.46)was annotated with the abiotic stress,and encoded for a calcium-dependent phospholipid binding protein(ANNEXIN). Therefore,we speculated that AhMYB44 maybe participate into regulation of peanut stress response process through intracellular calcium signal. This result might provide a reference for further research on the function and mechanism of AhMYB44.

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刘永惠,沈一,沈悦,等.利用酵母双杂交系统筛选花生AhMYB44互作蛋白[J].植物遗传资源学报,2020,21(1):201-207.

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  • 收稿日期:2019-10-29
  • 最后修改日期:2019-10-29
  • 录用日期:2019-11-07
  • 在线发布日期: 2020-01-17
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