核糖体失活蛋白(RIP)基因克隆及其表达载体的构建  

Cloning and Construction of Expression Vector of RIP Gene

作　　者：孙亚卿[1] 邵金旺[1] 张少英[1] 张洪霞[2] 

机构地区：[1]内蒙古农业大学甜菜生理研究所,内蒙古呼和浩特010018 [2]中国科学院上海植物生理生态研究所,上海200032

出　　处：《安徽农业科学》2007年第16期4770-4772,4803,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(30440048);内蒙古自然科学基金项目(200408020303)

摘　　要：以按玉米RIP基因(M83927)序列设计合成的特异性引物,用提取的玉米基因组DNA为模板,应用PCR技术扩增目的片段,测定并克隆玉米的RIP基因。序列分析表明,它们的核苷酸和氨基酸序列与GenBank中登录的CRIP(M83927)的同源性分别为99.90%和100%。RIP与植物表达载体pCAMBIA2301相连构建成为带有35S启动子和polyA终止子的植物表达载体p2301-RIP。The full length of RIP gene was amplified and cloned from maize by PCR.The DNA contained 953 bp with an open reading frame（ORF） of 921bp and encoded 307 amino acid residues.The deduced nucleotide acid sequence and amino acid sequence showed 99.90% and 100% homology to CRIP（M83927） respectively.The RIP gene was stickly inserted into BamHI and PstI site of the vector pCAMBIA2301 that was suitable for Agrobacterium-mediated DNA transformation.The new constructed plasmid p2301-RIP included GUS,nptII and RIP gene constructs.

关 键 词：RIP基因 DNA序列分析 克隆 载体构建 

分 类 号：Q786[生物学—分子生物学]