麻疯树pepc基因正义、反义植物表达载体构建与功能初步分析  

Constructing of Sense and Antisense pepc Expression Vector of Jatropha curcas and the Transformation

作　　者：范正琪[1] 卢孟柱[2] 李纪元[1] 田敏[1] 李辛雷[1] 吴开云[1] 陈东亮[1] 范妙华[1] 

机构地区：[1]中国林业科学研究院亚热带林业研究所,浙江富阳311400 [2]中国林业科学研究院林业研究所,北京100091

出　　处：《林业科学研究》2011年第2期165-170,共6页Forest Research

基　　金：浙江省重大科技专项重点项目"林源植物脂肪酸合成酶基因工程菌株的开发利用"(2006C12030)

摘　　要：根据麻疯树pepc基因和植物表达载体的酶切位点特征,分别将pepc基因全长序列3 000 bp正向插入pCAM-BIA2300,构建了正义表达载体pCAMBIA-Jcpepc,基因片段597 bp反向插入pBI121构建了反义表达载体pBI-Jcpepc。通过农杆菌介导,采用叶盘法转化烟草,通过对转基因植株的PCR和PCR-Southern检测,表明pepc基因已经正向、反向插入烟草基因组中。测定了转基因烟草的PEPCase酶活性,并通过脂肪酸的测定,转正义pepc基因烟草叶片的脂肪酸含量平均比对照降低44.3%,转反义pepc基因植株脂肪酸含量平均比对照增加26.3%。Antisense pepc gene expression vector was constructed by the 597 bp sequence of pepc gene ligated to pBI121 in the antisense orientation.And sense expression vector was recombined by the full-length（3 000 bp） sequence inserted to pCAMBIA2300.Mediated by Agrobacterium tumefaciens,the sense and antisense genes were transferred into the tobacco using leaf-disk method.Results of PCR and PCR-Southern determination indicated two gene sequences were integrated into the tobacco genome in the sense and antisense orientation.The activity of PEPCase was determined.It was found that the fat content of transgenic tobacco with sense pepc was 44.3 percent lower than that of control,and that with antisense pepc was 26.3 percent higher than that of the control.

关 键 词：麻疯树 PEPC基因 表达载体构建 转化 脂肪酸 

分 类 号：S718.46[农业科学—林学]