Acta Agrestia Sinica ›› 2015, Vol. 23 ›› Issue (3): 580-585.DOI: 10.11733/j.issn.1007-0435.2015.03.021

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Cloning and Transformation of MsMYB2 Gene from Medicago sativa L.

FENG Zi-rong1, SUN Yan1, CHAO Yue-hui2, YUAN Zhu1, LU Ji-xiao1   

  1. 1. Grassland Research Institute of China Agricultural University, Beijing 100093, China;
    2. The College of Forestry, Beijing Forestry University, Beijing 100083, China
  • Received:2014-05-21 Revised:2014-07-16 Online:2015-06-15 Published:2015-07-02

紫花苜蓿MsMYB2基因的克隆及转化

冯子蓉1, 孙彦1, 晁跃辉2, 袁柱1, 陆继肖1   

  1. 1. 中国农业大学草地研究所, 北京 100193;
    2. 北京林业大学林学院, 北京 100083
  • 通讯作者: 孙彦
  • 作者简介:冯子蓉(1990-),女,天津人,硕士研究生,从事牧草繁育和种子生产研究,E-mail: pipishan@yeah.net
  • 基金资助:

    国家牧草产业技术体系(CARS-35)专项;国家“十二五”科技支撑计划子课题“豆科及禾本科牧草良种扩繁技术研究与示范”专项(2011BAD17B01-02)资助

Abstract:

A gene named MsMYB2 was cloned from alfalfa by PCR technique with 834 bp coding domain sequence encoding 278 amino acids. The MsMYB2 gene was inserted into plasmid pBI121 by DNA recombination technology; and a plant expressing vector pBI-MsMYB2 was constructed. The plants of transgenic Arabidopsis thaliana with kanamycin resistance were screened through floral dip method and analyzed by PCR and RT-PCR. The results showed that the target gene was inserted into the genomes of Arabidopsis and expressed. Therefore, the transgenic Arabidopsis thaliana plants expressing MsMYB2 gene were successfully obtained in this study.

Key words: Medicago sativa L., Arabidopsis thaliana, MsMYB2 gene, Transformation

摘要:

为获得MsMYB2基因过量表达的转基因拟南芥(Arabidopsis thaliana)植株,利用PCR技术在紫花苜蓿(Medicago sativa)中克隆出MYB2基因并命名为MsMYB2,MsMYB2基因编码区全长834 bp,编码1条长度为278个氨基酸的多肽链。通过DNA重组技术将其与pBI121连接,成功构建了植物表达载体pBI-MsMYB2。通过花序侵染法获得具有卡那霉素抗性的转基因拟南芥植株,并通过PCR和RT-PCR对目的基因进行检测,结果证明目的基因已整合到拟南芥基因组中并且可以表达,成功获得了MsMYB2基因表达的拟南芥转基因植株。

关键词: 紫花苜蓿, MsMYB2基因, 拟南芥, 转化

CLC Number: