贝氏柯克斯体27kDa外膜蛋白基因的克隆与表达

目的　克隆和表达贝氏柯克斯体 (Coxiellaburnetii) 2 7kDa外膜蛋白基因 (com1 )。方法　采用PCR方法 ,从质粒pGEM -T/com1扩增出Com1基因片段 ,经鉴定后 ,将其克隆于原核表达载体 pQE30 ,构建成重组质粒 pQE30 /com1 ,IPTG诱导表达。结果　 (1 )获得长约 760bp的PCR片段 ,序列分析结果与已知com1序列相同。 (2 )SDS -PAGE检测表达产物 ,在相对分子量 2 7kDa处有表达条带。经薄层扫描分析 ,目的蛋白条带占全菌体蛋白的 1 7 2 %。 (3)经分析 ,有较好的抗原性。 (4)表达菌体超声破碎后 ,目的蛋白主要存在于包涵体中。结论　贝氏柯克斯体com1基因在大肠杆菌中获得了高效表达 ,所表达的重组蛋白能与Q热抗体发生反应

Cloning and expression of 27kDa outer membrane protein gene of Coxiella burnetii in E. Coli

Aim To clone and express the 27kDa outer membrane protein gene of Coxiella burnetii Methods com1 fragment was amplified from pGEM T/com1 by PCR,the fragment was cloned into prokaryotic expression vector pQE30;E coli cells were transformed with recombinant plasmid;the transformants were induced to express the fusion protein by IPTG Results The fusion protein from the transformants was approximate size of 27 kDa in SDS PAGE analysis and 17 2% of total cell protein in scanning analysis The fusion protein was recognized by serum from mice infected with immunoblot assay Conclusions The gene com1 can be efficiently expressed in E coli and the 27kDa fusion protein can react with the antibody to Coxiella burnetii