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植物研究 ›› 2014, Vol. 34 ›› Issue (4): 505-509.doi: 10.7525/j.issn.1673-5102.2014.04.013

• 论文 • 上一篇    下一篇

布鲁氏菌rOmp3148-74-BLS重组载体的构建与苜蓿的转化

赵亮1;王晶妍1,2*;王建英1,2;郭江波1,2;辛翠花1,2   

  1. 1.内蒙古科技大学数理与生物工程学院,包头 014010;2.内蒙古科技大学生物工程与技术研究所,包头 014010
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2014-07-20 发布日期:2014-07-20
  • 通讯作者: 王晶妍
  • 基金资助:
     

Construction of Brucella rOmp3148-74-BLS Expression Vector and Its Transformation into Alfalfa

ZHAO Liang;WANG Jing-Yan;*;WANG Jian-Ying;GUO Jiang-Bo;XIN Cui-Hua;   

  1. 1.School of Mathematics,Physics and Biological Engineering of Inner Mongolia University of Science and Technology,Baotou 014010;2.Institute of Bioengineering and Technology of Inner Mongolia University of Science and Technology,Baotou 014010
  • Received:1900-01-01 Revised:1900-01-01 Online:2014-07-20 Published:2014-07-20
  • Contact: WANG Jing-Yan
  • Supported by:
     

摘要: 将布鲁氏菌rOmp3148-74-BLS基因转入紫花苜蓿中苜一号,最终获得转基因苜蓿。本试验利用不依赖于连接反应的克隆(LigationIndependent Cloning,LIC)方法将布鲁氏菌rOmp3148-74-BLS基因克隆到植物表达载体pJG045上,并利用三亲本杂交的方法转化农杆菌AGL0。最后通过农杆菌介导法将该基因转化紫花苜蓿中苜一号。经PCR鉴定,筛选出转基因苜蓿植株。本试验为今后布鲁氏菌转基因植物疫苗的研发奠定了理论和实验基础。

关键词: 布鲁氏菌rOmp3148-74-BLS基因, 中苜一号, 转基因植物疫苗

Abstract: For the purpose of expression in plant an antigen of Brucella, the causative agent for zoonosis brucellosis, transgenic alfalfa was constructed using agrobacterium mediated gene transfer. Towards that end, a fusion gene rOmp3148-74-BLS containing Brucella Omp31 antigenic epitope spanning amino acids 48-74 and the lumazine synthase was cloned into vector pJG045 with ligation-independent cloning (LIC). The recombinant vector was transformed into Agrobacterium AGL0 by tri-parental hybrid method, which was used to infect calli of alfalfa Medicago sativa Zhongmu No.1. The transformed calli were identified by PCR, after stimulating the growth of roots, transgenic alfalfa was obtained. The experiment has laid experimental basis for the development of Brucella transgenic plants vaccine in the future.

Key words: Brucella rOmp3148-74-BLS, Medicago sativa Zhongmu No.1, transgenic plant vaccine

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