生物学杂志

生物学杂志

• 研究报告 • 上一篇    下一篇

水稻硝态氮响应的酵母双杂交文库的构建与分析

  

  1. 重庆师范大学 生命科学学院 植物环境适应分子生物学市重点实验室， 重庆 401331
  • 出版日期:2019-06-18 发布日期:2019-06-18
  • 通讯作者: 秦小健，博士，副教授，研究方向为植物分子遗传学，E-mail：qinxiaojian@cqnu.edu.cn
  • 作者简介:洪广成，硕士研究生，研究方向为生物化学与分子生物学，E-mail：625781104@qq.com
  • 基金资助:
    国家重点研发计划(2016YFD0100706);重庆师范大学人才启动基金(16XLB018);重庆市教委科学技术研究项目(KJ1600310)

Construction and analysis of a yeast two-hybrid library responding to nitrogen in rice （Oryza sativa）

  1. Chongqing Key Laboratory of Molecular Biology of Plants Environmental Adaptations，College of Life Sciences, Chongqing Normal University, Chongqing 401331, China
  • Online:2019-06-18 Published:2019-06-18

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摘要：  酵母双杂交是在真核生物酵母细胞体内研究蛋白质之间相互作用的一种高效分子生物学技术。为了研究水稻氮吸收与利用的分子调控机理，以硝态氮(5 mmol/L KNO3)诱导处理的水稻 9311幼苗为材料，构建水稻在高氮响应条件下的酵母双杂交cDNA文库。实验结果表明，构建的氮响应文库滴度为1.7×108 cfu/mL，文库的重组率为 95% ，插入片段大小在 500~1500 bp之间。从文库中随机挑选10个单克隆，测序后通过NCBI比对分析结果显示9个克隆可以从水稻基因库中找到对应序列。文库的构建将为水稻氮素吸收与利用的分子调控机理研究提供帮助。

关键词: 水稻, 氮吸收与利用, 酵母双杂交, 文库构建

Abstract: Yeast two hybrid is an efficient molecular biology technique to study the interaction of proteins in eukaryotic yeast cells. In order to study the molecular regulation mechanism of nitrogen uptake and utilization in rice, we constructed a yeast two hybrid cDNA library in response to high nitrogen conditions by using nitrate (5 mmol/L KNO3) induced rice 9311 seedlings as materials. The test results showed that the titer of the library is 1.7×108 cfu/mL, the recombination rate of the library is 95% and the insert fragment size is 500-1500 bp. Furthermore, 10 clones were randomly selected from the library and followed sequencing, the results of NCBI alignment analysis showed that 9 clones could find the corresponding sequences from the rice gene pool. Summarily, the construction of this library helps to study the gene regulation of nitrogen absorption and utilization in rice.

Key words: rice, nitrogen absorption and utilization, yeast two hybrid, library construction

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