红斑丹毒丝菌果糖二磷酸醛缩酶基因的克隆和表达  

Cloning and Expression of the Fructose-Bisphosphate Aldolase Gene of Erysipelothrix rhusiopathiae

作　　者：恩特马克.布拉提白 张娜[1] 吾鲁木汗.那孜尔别克 

机构地区：[1]伊犁师范学院生物与地理科学学院,新疆伊宁835000

出　　处：《伊犁师范学院学报（自然科学版）》2016年第4期41-45,共5页Journal of Yili Normal University：Natural Science Edition

基　　金：国家自然科学基金项目(31560702;31360613)

摘　　要：用PCR技术从红斑丹毒丝菌C43065株基因组DNA中扩增出编码果糖二磷酸醛缩酶(fructose-bisphosphate aldolase)的fba基因序列,构建原核表达载体p QE30-fba,转化大肠埃希杆菌M15,用IPTG诱导表达目的蛋白.结果显示:红斑丹毒丝菌C43065株fba基因ORF大小为867 bp,编码288个氨基酸,与参考株序列相似性为99%;在大肠埃希杆菌M15中获得分子量约为32 k Da的表达产物,与抗His标签单克隆抗体发生特异性反应.本研究获得的重组蛋白FBA,为进一步开展FBA免疫功能和致病性研究奠定了基础.The fba gene encoding the fructose-bisphosphate aldolase （FBA） was amplified by PCR from the genomic DNA of Erysipelothrix rhusiopathiae strain C43065, then cloned into the prokaryotic expression vector pQE30 and expressed in E. coli M15 by IPTG induction. The expressed protein was identified by SDS-PAGE and Western blotting. The open reading frame of the fba gene was 867 bp in length, and the nucleotide sequence of the fba gene from the C43065 was 99% similar to that of the fba gene in the previously reported E. rhusiopathaie strain Fujisawa. A single protein band with a molecular weigth of 32 kDa was expressed in E. coli M15, and the protein was recognized specifically by an anti His-Tag mouse monoclonal antibody. The recombinant protein FBA can be used for further study of the protective activity and pathogenesis of E. Rhusiopathiae.

关 键 词：红斑丹毒丝菌 果糖二磷酸醛缩酶 原核表达 

分 类 号：Q786[生物学—分子生物学]