安徽农业科学2019，Vol.47Issue(2)：96-98,3.DOI:10.3969/j.issn.0517-6611.2019.02.028
拟南芥MDH2基因GFP载体构建及转基因植株的筛选鉴定
Construction of Arabidopsis MDH2-GFP Vector and Identification of Transgenic Lines
摘要
Abstract
[Objective] In order to further study the function of MDH2 gene in response to cadmium stress in plant, the vector of Arabidopsis MDH2-GFP was built up and transgenic lines were constructed.[Method] The RNA of wild-type Arabidopsis was extracted and reverse transcribed into cDNA, the full-length MDH2 coding sequence was amplified by PCR. The amplified MDH2 gene and pXB94-GFP plasmid were digested and connected by double enzymes.The connected product was transformed into E.coli competent cells, then the true recombinant plasmid was transformed into Agrobacterium tumefaciens competent cells and the positive single colony was identified by colony PCR, then it was transferred into wild-type Arabidopsis by the floral-dip method.Finally, the positive transgenic plants were obtained by resistance screening and PCR identification.[Result]MDH2 gene was successfully cloned and the positive transgenic lines were obtained by selective medium with resistance.[Conclusion]The successful acquisition of MDH2-GFP transgenic lines laid the foundation for further research on the function of MDH2 gene.关键词
拟南芥/MDH2/载体/转基因植株Key words
Arabidopsis/MDH2/Vector/Transgenic lines分类
生物科学引用本文复制引用
王媛媛,韩洋洋,耿庆鎏,朱香豫,曹树青,樊婷婷..拟南芥MDH2基因GFP载体构建及转基因植株的筛选鉴定[J].安徽农业科学,2019,47(2):96-98,3.基金项目
中央高校基本科研业务费专项资金资助(JZ2018HGTB0248) （JZ2018HGTB0248）
