水稻核糖体蛋白基因OsRPL2的克隆、序列分析及表达载体构建  

Cloning and Sequence Analysis, Expression Vector Construction of a Ribosomal Protein Gene Os RPL2 from Rice

作　　者：宋丹妮[1] 王春台[1] 刘学群[1] 谭艳平[1] 刘新琼[1] 程钢[1] 

机构地区：[1]中南民族大学/武陵山区特色资源植物种质保护与利用湖北省重点实验室,武汉430074

出　　处：《湖北农业科学》2015年第11期2773-2777,共5页Hubei Agricultural Sciences

基　　金：国家自然基金项目(31170296);湖北省自然科学基金项目(2012FFB074)

摘　　要：依据鉴定得到的水稻(Oryza sativa L.)核糖体蛋白质Os RPL2的序列设计了引物,从水稻叶片的c DNA中扩增得到目的片段。并对目的基因序列进行了测序鉴定和序列分析,同时在大肠杆菌中构建水稻核糖体蛋白质基因Os RPL2的原核表达载体p GEX-4T1-RPL2,转化入BL21(DE3)后利用IPTG诱导外源蛋白质表达并检测。结果表明,成功扩增出了水稻核糖体蛋白质基因Os RPL2的c DNA序列,大小为1 570 bp,其编码的蛋白质含有502个氨基酸。根据序列分析的结果,水稻Os RPL2基因与多个植物核糖体蛋白质L2基因具有较高的相似性,并且其编码的Os RPL2蛋白质具有2个与核糖体蛋白质功能相关的功能域。在此基础上,构建了具有p GEX-4T1-RPL2重组子,诱导表达后,纯化得到了带有GST标签的可溶性Os RPL2蛋白质。On the basis of identified rice ribosomal protein sequences,a primer was designed to amplify OsRPL2 gene from rice （Oryza sativa L.） leaves＇ cDNA. The obtained sequence was sequenced and analyzed, meanwhile,inserted into plasmidp GEX-4T1 to generate a prokaryotic expression vector pGEX-4T1-RPL2 which was transferred into the Escherichia coli BL21 （DE3） strain, and then the protein expression was detected after induced by IPTG. The results showed that, the amplified fragment was identical to the cDNA sequence of OsRPL2 and contained a complete open reading frame of 1 570 bp, encoding a protein of 502 amino acid residues. Meanwhile, OsRPL2 gene has a high similarity with several plant ribosomal L2 genes, and the encoded OsRPL2 protein has two domains associated with ribosomal protein function. In addition, the soluble OsRPL2 protein with GST label was purified after IPTG inducing.

关 键 词：水稻(Oryza SATIVA L.) 核糖体蛋白质基因 原核表达 

分 类 号：Q781[生物学—分子生物学]