嗜肺军团菌momp原核重组质粒的构建和表达  

The Construction and Expression of Recombinant Plasmid pET-momp of Legionella Pneumophila Major Outer Membrane Protein Gene

作　　者：窦娇莹[1] 曹秀琴[2] 杨志伟[1] 

机构地区：[1]宁夏医科大学基础医学院病原生物学与免疫学系,银川750004 [2]宁夏医科大学生育力保持省部共建教育部重点实验室,银川750004

出　　处：《宁夏医科大学学报》2011年第11期1016-1018,1022,共4页Journal of Ningxia Medical University

基　　金：国家自然科学基金(30860264);宁夏自然科学基金(NZ0992)

摘　　要：目的构建嗜肺军团菌momp原核重组质粒,并纯化重组蛋白。方法以嗜肺军团菌LP1型DNA为模板,PCR扩增得到momp基因,定向克隆至原核载体PET32a(+)中,经酶切及测序鉴定正确后,转化入大肠杆菌BL21中,用IPTG诱导并用SDS-PAGE电泳进行分析,其产物用亲和层析法进行纯化。结果扩增出831bp的momp基因,构建重组质粒pET-momp,诱导表达及纯化出50KD的蛋白。结论成功构建momp基因的原核表达载体并得到高效表达。Objective To construct recombinant plasmid pET-momp of Legionella Pneumophila major outer membrane protein Gene and to purify the recombinant protein.Methods LP1 type of Legionella pneumophila DNA（a template momp gene） was amplified by PCR and cloned into prokaryotic vector PET32a（＋）.After restriction analysis and DNA sequencing,It was transformed into E.coli BL21,then it was induced with IPTG and analyzed by SDS-PAGE.Its products were purified by affinity chromatography.Results 831bp momp gene was amplified and,the recombinant plasmid pET-momp was constructed.50kD MOMP protein was successfully purified.Conclusion Momp gene was successfully constructed and highly expressed.

关 键 词：嗜肺军团 momp基因 表达 纯化 

分 类 号：R378[医药卫生—病原生物学]