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J. Hefei Univ. Tech. (Nat. Sci.)

合肥工业大学学报(自然科学版)

JOURNAL OF HEFEI UNIVERSITY OF TECHNOLOGY (NATURAL SCIENCE)

Competent Authorities: Ministry of Education of the People's Republic of China

Sponsor by: Hefei University of Technology

ISSN 1000-1646, CN 34-1001/N

Construction and cellular localization of CTM2-GFP transgenic Arabidopsis thaliana

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CTM2-GFP 转基因拟南芥构建及细胞定位研究

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Oct.2024, Vol.47 No.10: 1377-1380, 1411

DOI: 10.3969/j.issn.1003-5060.2024.10.013

Authors

LI Yaping, CAO Lei, ZHENG Pengpeng, FAN Tingting

(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China)

Abstract and Keywords

Abstract: The mutant plants with cadmium(Cd) tolerance obtained by screening the mutant library may be involved in the regulation of plant Cd stress tolerance. In this paper, pART27-CTM2-GFP vector was constructed by homologous recombination method, and the recombinant plasmid of pART27-CTM2-GFP was transferred into Agrobacterium GV3101 and then into the wild-type Arabidopsis plant by floral dip transformation. CTM2-GFP transgenic positive plants were obtained through resistance screening and polymerase chain reaction (PCR) identification, and homozygous transgenic plants were obtained through resistance isolation and identification. Finally, CTM2 gene expression was detected by qPCR to screen the best CTM2-GFP transgenic lines. In addition, by observing the green fluorescent protein (GFP) fluorescence in CTM2-GFP transgenic plants, it was found that CTM2 was a cell membrane localization protein. The results provide genetic material support for the in-depth study of the role of CTM2 gene in regulating the molecular mechanism of plant Cd tolerance.

Keywords: Arabidopsis thaliana; CTM2 gene; plant expression vector; transgenic plant; cadmium tolerance

作者信息

李亚平，曹蕾，郑朋朋，樊婷婷

（合肥工业大学食品与生物工程学院，安徽合肥230601）

摘要和关键词

摘要: 通过筛选突变体库发现的镉耐受突变体植株可能参与了植物Cd胁迫的调控。文章通过同源重组方法构建pART27-CTM2-GFP载体，并将pART27-CTM2-GFP重组质粒转入农杆菌GV3101，采用浸花转染法转入野生型拟南芥植株；通过抗性筛选和聚合酶链式反应（polymerase chain reaction，PCR）鉴定获得CTM2-GFP转基因阳性植株，进行抗性分离鉴定获得纯合转基因植株；最后通过qPCR检测CTM2基因表达，筛选最佳的CTM2-GFP转基因株系；通过观察CTM2-GFP转基因植株中绿色荧光蛋白（green fluorescent protein，GFP）的荧光发现CTM2为细胞膜定位蛋白。研究结果为深入研究CTM2基因在调控植物镉耐受分子机制中的作用提供了遗传材料支撑。

关键词: 拟南芥；CTM2基因；植物表达载体；转基因植株；镉耐受

Funding

安徽省自然科学基金资助项目（1508085QC50）

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