安徽农业科学2019，Vol.47Issue(2)：86-88,3.DOI:10.3969/j.issn.0517-6611.2019.02.025
拟南芥SPM12基因GFP载体构建及转基因植株筛选
Construction of Arabidopsis SPM12-GFP Vector and Identification of Transgenic Lines
摘要
Abstract
[Objective] To further study the function of the SPM12 gene, the SPM12 gene GFP vector and its transgenic plants were constructed by wild-type Arabidopsis thaliana with genetic background in Columbia (Columbia, Col). [Method] The full-length CDS sequence of SPM12 gene was amplified by PCR from the cDNA template which was reverse transcription from the RNA of wild-type Arabidopsis thaliana, and ligated it with the plasmid of GFP vector; then transformed the recombinant plasmid which was successfully constructed into the cells of E. coli DH5α, picked the positive colonies and identified it by PCR, and then transformed its plasmid into the competent cells of Agrobacterium GV3101, the positive colonies were identified by PCR, then it was transferred into wild-type Arabidopsis thaliana through inflorescence-infiltrated method. After receiving the seeds, the positive plants were screened by selective medium with resistance. [Result] The CDS fragment of SPM12 gene and SPM12-GFP recombinant plasmid was obtained, the vector of SPM12-GFP and its transgenic plant was got. [Conclusion] The transgenic plants SPM12-GFP in Arabidopsis thaliana were successfully obtained, which laid a foundation for further study of the function and molecular mechanism of SPM12 gene in Arabidopsis thaliana.关键词
拟南芥/SPM12/载体/GFP转基因植株Key words
Arabidopsis thaliana/SPM12/Vector/GFP transgenic lines分类
生物科学引用本文复制引用
方雪,徐洁娜,孟杏楠,曹树青,樊婷婷..拟南芥SPM12基因GFP载体构建及转基因植株筛选[J].安徽农业科学,2019,47(2):86-88,3.基金项目
中央高校基本科研业务费专项资金项目(JZ2018HGTB0248) （JZ2018HGTB0248）
