金黄色葡萄球菌isdB基因克隆、表达及其抗原性鉴定  

Prokaryotic Expression of Staphylococcus aureus IsdB and Identification of Its Antigenicity

作　　者：崔莉[1,2] 朱战波[1] 朱洪伟[1] 崔玉东[3] 朴范泽[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]广东温氏食品集团有限公司江苏分公司 [3]黑龙江八一农垦大学生命科技学院

出　　处：《河北科技师范学院学报》2011年第1期16-20,共5页Journal of Hebei Normal University of Science & Technology

基　　金：黑龙江省科技厅重大项目(项目编号:GA09B301-2)

摘　　要：为了构建含牛源金黄色葡萄球菌isdB基因的原核表达载体,并确定其在大肠杆菌表达系统中的表达效果,应用PCR方法扩增出isdB基因片段与原核表达载体pQE-30,构建了重组原核表达载体pQE-30-isdB,将该重组载体转化至E.coliXL1-Blue中诱导表达蛋白。经SDS-PAGE和Western blot鉴定,PCR扩增出约1 938 bp的基因片段,测序结果与GenBank上发表的MW2株核苷酸和氨基酸序列的一致性均为98.6%;成功表达出约72 ku的蛋白,与IsdB蛋白的相对分子质量大小一致;Western blot分析表明,该蛋白具有金黄色葡萄球菌的抗原性。本次试验成功构建了IsdB蛋白的原核表达系统。To construct the prokaryotic expression vector of Staphylococcus aureus isdB gene from cow and express the gene in E.coli.,isdB gene was amplified by PCR,to expression vector pQE-30-isdB,the prokaryotic expression vector pQE-30-isdB was constructed and transformed to E.Coli XL1-Blue for expression.SDS-PAGE and Western Blot showed that a gene fragment at a length of 1 938 bp was amplified.Sequencing results showed that the isdB gene of the isolated strain shares 98.6% homology in nucleotide and amino acids sequence with that of the S.aureus strain MW2 in GenBank.The protein with a relative molecular weight of 72 ku was expressed.Western-blotting analysis indicated that the protein had antigenic activity of IsdB.This experiment successfully constructed the IsdB protein＇s prokaryotic expression system.

关 键 词：金黄色葡萄球菌 ISDB 原核表达 

分 类 号：S855.11[农业科学—临床兽医学]