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作物学报 ›› 2010, Vol. 36 ›› Issue (2): 341-346.doi: 10.3724/SP.J.1006.2010.00341

• 研究简报 • 上一篇    下一篇

花生白藜芦醇合成酶基因PNRS1的克隆及其在原核中的表达

韩晶晶1,2,刘炜1,*,毕玉平1,2   

  1. 1 山东省农业科学院高新技术研究中心 / 农业部黄淮海作物遗传改良与生物技术重点开放实验室，山东济南 250100；2 山东师范大学生命科学学院，山东济南 250014
  • 收稿日期:2009-11-06 修回日期:2009-10-04 出版日期:2010-02-10 网络出版日期:2009-12-21
  • 基金资助:
    本研究由转基因生物新品种培育科技重大专项(2009ZX08001-010B)和山东省农业科学院博士科研启动基金项目(2007YBS008)资助。

Molecular Cloning of Peanut Resveratrol Synthetic Enzyme 1(PNRS1) and its Expression in Prokaryote

HAN Jing-Jing1,2,LIU Wei1,*, BI Yu-Ping1,2
  

  1. 1 Hi-Tech Research Center, Shandong Academy of Agricultural Sciences / Key Laboratory of Crop Genetic Improvement and Biotechnology, Huanghuaihai, Ministry of Agriculture, Ji’nan 250100 China; 2 College of Life Sciences, Shandong Normal University, Ji’nan 250014, China
  • Received:2009-11-06 Revised:2009-10-04 Published:2010-02-10 Published online:2009-12-21

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2053

被引次数

11

摘要：

采用RT-PCR克隆花生白藜芦醇合成酶(resveratrol synthetic enzyme, RS)基因全长，命名为PNRS1，GenBank登录号为FM955393。序列分析表明该基因的开放读码框1 170 bp，编码389个氨基酸残基。以花生品种鲁花14基因组DNA为模板经PCR扩增，获得该基因的基因组序列长1 537 bp，包含2个外显子和1个内含子。比较发现，PNRS1与其他已知RS序列的同源性达91%~95%。表达模式分析显示，PNRS1在花生根中特异表达，并可被紫外线UV-B诱导。PNRS1与pET-28a(+)构建原核表达载体，经IPTG诱导后可表达获得相对分子量约为46 kD的外源融合蛋白。以上结果证实PNRS1属花生RS基因家族成员，并为进一步分析该基因的功能奠定了基础。

关键词: 花生, 白藜芦醇合成酶, 表达模式, 原核表达, 融合蛋白

Abstract:

Resveratrol synthetic enzyme (RS) is a key and necessary enzyme that plays important role in resveratrol synthesis pathway. To uncover and characterize the function of the RS in plant development process, we isolated a RS gene PNRS1 (FM955393) by RT-PCR using total RNA of peanut seed as template, and the gene PNRS1 was expressed in E. coli prokaryote for further analysis. The results showed that the PNRS1 had a 1 170 bp open reading frame encoding 389 amino acid polypeptide, and exhibiting high similarities with other members of RS genes family. Expression pattern analysis indicated that the PNRS1 was specially expressed in peanut root, and could be induced by UV-B treatment. The recombinant PNRS1 protein product with the molecular weight about 46 kD could also be detected in E. coli protein expression system. These results suggested that the PNRS1 was a mumber of RS family with peculiar expression patterns compared with other ones. As protein is the functional executor of a gene, the protein product of PNRS1 finally will take effect in the later processes of plant development, and shed light on the stress-resistant breeding and cultivation.

Key words: Peamut, Resveratrol synthetic enzyme(RS), Expression pattern, Prokaryotic expression system, Fusion protein




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