与水稻黑条矮缩病毒p5b互作的水稻基因片段筛选

doi:10.3969/j.issn.10017216.2012.05.004

中国水稻科学 ›› 2012, Vol. 26 ›› Issue (5): 537-541.DOI: 10.3969/j.issn.10017216.2012.05.004

与水稻黑条矮缩病毒p5b互作的水稻基因片段筛选

陆颖^1，2,羊健² ,张恒木² ,陈剑平^2，*

¹ 南京农业大学植物保护学院，江苏南京 210095； ² 浙江省农业科学院病毒学与生物技术研究所/浙江省植物有害生物防控国家重点实验室培育基地/农业部植保生物技术重点开放实验室，浙江杭州 310021；

收稿日期:2012-01-16 修回日期:2012-03-02 出版日期:2012-09-10 发布日期:2012-09-10
通讯作者: 陈剑平2，*
基金资助:
国家973计划资助项目（2010CB1Z6203);浙江省自然科学基金资助项目（Y3090657）；国家科技支撑计划资助项目（2012BAD19B03)。

Screening of Rice Genes Interacted with p5b of Rice blackstreaked dwarf virus

LU Ying ^1,2, YANG Jian ², ZHANG Hengmu ², CHEN Jianping ^2,*

¹ College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; ²State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control /Key Laboratory of Plant Protection and Biotechnology, Ministry of Agriculture / Zhejiang Provincial Key Laboratory of Plant Virology /Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;

Received:2012-01-16 Revised:2012-03-02 Online:2012-09-10 Published:2012-09-10
Contact: CHEN Jianping2,*

摘要/Abstract

摘要： 由水稻黑条矮缩病毒基因组S5片段第2个开放阅读框（ORF）编码的p5b是一个功能未知的病毒非结构蛋白。构建了酵母双杂交体系的含水稻cDNA表达文库的pGADT7载体，同时将p5b 基因构建到诱饵质粒pGBKT7上（pGBKp5b），Western Blot和自激活实验结果显示p5b可在酵母中正确表达且无自激活活性。进一步利用p5b蛋白作为诱饵用酵母双杂交筛选水稻cDNA表达文库，鉴定出一些参与光合作用、呼吸作用等重要代谢过程的关键酶基因片段，这些酶可能与p5b蛋白之间存在相互作用，推测这些互作在病株症状发展过程中起作用。

关键词: 水稻黑条矮缩病毒, 互作蛋白, 酵母双杂交

Abstract: The second open reading frame on the genome segment 5 (S5) of Rice blackstreaked dwarf virus (RBSDV) encodes a viral nonstructural protein (p5b) with unknown function. To reveal the function of p5b, its gene was ligated into bait plasmid pGBKT7 and an expression library containing rice cDNAs was constructed using plasmid pGADT7 of yeast twohybrid. The bait protein p5b was detected in yeast by western blot and the result of an autoactivation test showed that p5b could not autonomously activate the expression of reporter genes in yeast. Then the bait protein p5b was used for screening the expression cDNA library of rice plant. Gene fragments of some pivotal enzymes, involved in photosynthesis, respiration, and other important metabolism processes, were identified to interact with p5b in yeast, suggesting that these interactions may play roles in symptom development of infected plants.

Key words: Rice blackstreaked dwarf virus (RBSDV), interaction protein, yeast twohybrid (Y2H)

中图分类号:

陆颖1，2,羊健2 ,张恒木2 ,陈剑平2，*. 与水稻黑条矮缩病毒p5b互作的水稻基因片段筛选[J]. 中国水稻科学, 2012, 26(5): 537-541.

LU Ying1,2, YANG Jian2, ZHANG Hengmu2, CHEN Jianping2,*. Screening of Rice Genes Interacted with p5b of Rice blackstreaked dwarf virus[J]. Chinese Journal of Rice Science, 2012, 26(5): 537-541.

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与水稻黑条矮缩病毒p5b互作的水稻基因片段筛选

Screening of Rice Genes Interacted with p5b of Rice blackstreaked dwarf virus

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