草地学报

草地学报 ›› 2016, Vol. 24 ›› Issue (6): 1388-1392.DOI: 10.11733/j.issn.1007-0435.2016.06.035

• 研究专报 • 上一篇    

紫花苜蓿MsHST基因克隆及拟南芥转化

李奕稹1, 晁跃辉2, 康俊梅2, 杨青川2, 金洪1,2   

  1. 1. 内蒙古农业大学农学院, 内蒙古 呼和浩特 010019;
    2. 中国农业科学院北京畜牧兽医研究所, 北京 100193
  • 收稿日期:2015-03-31 修回日期:2015-10-31 出版日期:2016-12-15 发布日期:2017-03-18
  • 通讯作者: 金洪
  • 作者简介:李奕稹(1987-),女,内蒙古四子王旗人,硕士研究生,研究方向为植物资源保护与利用,E-mail:15661065466@163.com
  • 基金资助:

    十二五国家科技支撑子课题“紫花苜蓿品质性状相关基因的挖掘与功能分析”（2014CB138703-2）；中国农业科学院北京畜牧兽医研究所基本科研业务费项目（2014ywf-zd-2）资助

Cloning of MsHST Gene from Alfalfa and Obtainment of Transgenic Arabidopsis thaliana

LI Yi-zhen1, CHAO Yue-hui2, KANG Jun-mei2, YANG Qing-chuan2, JIN Hong1,2   

  1. 1. College of Ecology and Environmental Science, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia 010019, China;
    2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2015-03-31 Revised:2015-10-31 Online:2016-12-15 Published:2017-03-18

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摘要：

运用RT-PCR法从紫花苜蓿（Medicago sativa）中成功克隆出长度约1100 bp的尿黑酸法呢基转移酶基因MsHST，采用qRT-PCR的方法，分析MsHST基因的表达特征，结果显示MsHST在所有组织中均有表达，但在叶片中表达量最高，根中最低；随叶片的衰老，表达量降低。且构建了超表达载体pBI-MsHST，并获得拟南芥（Arabidopsis thaliana）转基因植株，经PCR，RT-PCR和GUS染色检测转基因植株，初步证实目的基因在拟南芥中表达。

关键词: 紫花苜蓿, MsHST, 拟南芥, 转基因

Abstract:

By using RT-PCR, we cloned a length of 1100 bp of urine black acid base transferase gene from alfalfa (Medicago sativa). It was named as MsHST, and its expression characteristics were analyzed with qRT-PCR. The results showed that MsHST in all the tested organs, including root, stem, leaf and flower were expressed, but its expression in blade was the highest, expression in root was the lowest. The gene expression was decreased with the senescence of leaf. Using recombinant DNA technology, the MsHST was inserted to expression vector pBI121 to make a construct ofpBI-MsHST. Through Agrobacterium mediated-transfermation, putative transgenic Arabidopsis(Arabidopsis thaliana)plants were obtained, and subjected to PCR, RT-PCR and GUS staining assay, the results showed that the target gene has been integrated to the Arabidopsis thaliana genome and was expressed.

Key words: Alfalfa, MsHST, Arabidopsis thaliana, Transgenic

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