AtCS82基因T-DNA插入突变体筛选和过表达转基因植株的获得  

Screening of T–DNA insertion mutants and construction of plants with AtCS82 over expressed

作　　者：陈锦华[1] 郭磊[2] 易力雄[1] 刘春林[2] 阮颖[1] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128 [2]湖南农业大学农学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2013年第3期253-258,共6页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31071455)

摘　　要：AtCS82是拟南芥中一个功能未知的基因,推测该基因可能与种子油脂合成代谢相关。为探明AtCS82基因的功能,采用半定量RT-PCR法对AtCS82基因在拟南芥不同组织(茎、叶、花、果荚)中的表达情况进行了分析,发现其在果荚中的表达量较高,而在茎、叶、花中的表达量明显偏低,推测AtCS82基因可能参与种子发育;使用三引物法对T-DNA插入突变体进行纯合子筛选,并通过RT-PCR检测纯合子植株中AtCS82基因的表达情况,试验结果证明获得了稳定遗传的AtCS82基因沉默纯合突变体;通过构建pCAMBIA1300-35S::CS82过表达载体,并转化拟南芥哥伦比亚野生型(Col)的方式,获得了AtCS82基因的过表达转基因植株。The function ofAtCS82 gene in Arabidopsis is still unknown, but it is supposed to be associated with seed oil anabolism. To investigate the function of ArCS82 gene, semi-quantitative RT-PCR was used to analyze the expression of ARCS82 gene in different tissues of Arabidopsis （stem, leaf, flower, pod）, the results show expression of ArCS82 gene was high in pod, but low in stem, leaf and flower, indicating the participation of ARCS82 gene in seed development. Three primers were used to screen homozygous mutants with T-DNA insertion, and the expression of ARCS82 gene in homozygous mutant plant was tested using RT-PCR, the results proved the obtaining of stable pure mutant with AtCS82 gene silenced. AtCS82-overexpressing transgenic plants were obtained by constructing pCAMBIA1300-35S∷CS82 recombinant plasmid, followed by transforming into Arabidopsis thaliana of Columbia ecotype （Col）.

关 键 词：拟南芥 AtCS82基因 纯合子筛选 过表达 转基因植株 

分 类 号：Q786[生物学—分子生物学]