巴尔通体HBPC蛋白原核重组表达及抗原性研究  

Expression and Antigenic Investigation of Recombinant HBPC Protein of Bartonella tribocorum

作　　者：叶曦[1] 伍小春[2] 李国伟[1] 罗炜[1] 王团老[2] 

机构地区：[1]厦门市海沧区疾病预防控制中心,福建厦门361026 [2]厦门大学药学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2013年第3期405-410,共6页Journal of Xiamen University：Natural Science

基　　金：厦门市科技计划项目(WZK-0629)

摘　　要：为了寻找可用于巴尔通体(Bartonella)检测的特异性抗原,利用PCR方法从巴尔通体厦门分离株(B.tribocorum_XM)的基因组DNA中扩增出血红素结合蛋白C(HBPC)蛋白基因,并将该基因的编码区克隆到pGEX-4T-1原核表达载体中,构建带GST标签的融合蛋白重组表达载体,并转化大肠杆菌(Escherichia coli DH5α),诱导表达GST-HBPC,通过亲和层析技术纯化GST-HBPC.最后运用蛋白质斑点印迹试验(Western blot)和间接ELISA法检验GST-HBPC是否具有抗原性.结果表明,所构建的表达载体可在大肠杆菌中大量表达GST-HBPC,Western blot和间接ELISA显示GST-HBPC能够特异性地与感染动物血液样本发生免疫反应.因此,体外重组的巴尔通体抗原蛋白HBPC可作为潜在的抗原用于巴尔通体血清学免疫检测.Here we tried to find specific antigen for Bartonella deteclion. HBPC gene was amplified from the genomic DNA of B. tri- bocorum isolated from Xiamen region through PCR assay,and sub-cloned into the prokaryotic expression vector pGEX-4T-1 vector to generate the construct for expressing GST-fusion protein GST-HBPC, the vector was transformed into Escherichia coli DH5α for in duction to express GST-HBPC. GST-HBPC was purified through affinity chromatography assay. The antigenicity of GST HBPC was investigated with Western blot experiment and indirect ELISA assay. The results demonstrated that we successfully generated the prokaryotic expression vector for recombinant HBPC protein. The expression vector can be induced to produce GST-HBPC efficient ly,and the Western blot experiment and ELISA assay indicated that GST-HBPC immunologically and specifically reacted with the blood samples from the infected animal. Therefore,the expressed recombinant Bartonella HBPC protein is a potential antigen of Bar tonella. The results are potentially useful for serological detection of Bartonella,

关 键 词：巴尔通体 HBPC 抗原 

分 类 号：Q782[生物学—分子生物学]