荠菜雌蕊调控转录因子SPT基因的克隆与拟南芥遗传转化  

Cloning of the Gene of Pistil-regulated Transcription Factor in Capsella bursa-pastoris and Its Genetic Transformation in Arabidopsis thaliana

作　　者：张莉芳 杨正修[1] 张学文[1] 龙炎杏 赵燕[1] ZHANG Lifang;YANG Zhengxiu;ZHANG Xuewen;LONG Yanxing;ZHAO Yan(College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha,Hunan 410128, China)

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128

出　　处：《作物研究》2018年第3期202-207,共6页Crop Research

基　　金：湖南省教育厅科学研究项目(15K060)

摘　　要：根据Gen Bank收录的拟南芥At SPT基因c DNA序列设计引物,采用同源克隆的方法从野生型荠菜(Capsella bursa-pastoris)中RT-PCR扩增出荠菜Cb SPT基因全长c DNA,通过生物信息学分析表明:荠菜Cb SPT基因c DNA序列1191 bp,其中开放阅读框1047 bp,编码348个氨基酸,与拟南芥At SPT基因的核酸序列和氨基酸序列的同源性分别达到98%和93%左右,可以判断为荠菜Cb SPT基因c DNA序列(Cb SPT)。以Ti质粒p WM101为基础,构建了由Ca MV35S启动子调控的Cb SPT基因植物表达载体p WM101-35S::Cb SPT。采用根癌农杆菌介导的浸花序法转化荠菜的同科植物拟南芥,获得了转Cb SPT基因的拟南芥植株。转基因拟南芥心皮果荚形态大小发生了一定的变化,说明荠菜Cb SPT基因在拟南芥中的组成型表达对拟南芥心皮形态和大小都产生了一定影响,但并没有使拟南芥表现出荠菜短角果的形态。The primers were designed according to the Arabidopsis At SPT c DNA sequence contained in Gen Bank,with the method of Homology-based cloning,the full-length c DNA of the Cb SPT gene was amplified by RT-PCR in Capsella bursa-pastoris. Bioinformatics analysis showed that： the capsella bursa-pastoris Cb SPT gene c DNA sequence 1191 bp,with 1047 bp open reading frames,coding 348 amino acids,the homology of nucleic acid sequence and amino acid sequence of the At SPT gene of Arabidopsis was 98% and 93% respectively,which could be judged as the Cb SPT gene c DNA sequence（ Cb SPT）. Based on Ti plasmid p WM101,Cb SPT gene plant expression vector p WM101-35 S： ： Cb SPT regulated by Ca MV35 S promoter was constructed. Arabidopsis thaliana was transformed by the method of agrobacterium,Arabidopsis plants transformed with Cb SPT gene were obtained. There were some changes in the shape and size of the transgenic Arabidopsis carpel pods,which means that the expression of the Cb SPT gene in Arabidopsis thaliana had a certain effect on the shape and size of Arabidopsis thaliana. But,it did not change the Arabidopsis into the shape of Capsella bursa-pastoris.

关 键 词：荠菜 SPT基因 克隆 遗传转化 

分 类 号：Q785[生物学—分子生物学]