幽门螺杆菌hspA-ureB融合基因在乳酸菌中的表达  

Fusion expression of Helicobacter pylori hspA-ureB genes in lactic acid bacterium

作　　者：张荣光[1,2] 孙楠[1] 段广才[1,2] 彭晓燕[1] 陈帅印[1] 范清堂[2,3] 

机构地区：[1]郑州大学公共卫生学院流行病与统计学教研室,河南郑州450001 [2]河南省分子诊断与医学检验技术协同创新中心(新乡医学院),河南新乡453000 [3]河南省分子医学重点实验室,河南郑州450001

出　　处：《中国卫生检验杂志》2016年第7期921-923,928,共4页Chinese Journal of Health Laboratory Technology

基　　金：中国博士后科学基金特别资助项目(200801273);河南省分子诊断与医学检验技术协同创新中心(新乡医学院)资助项目(XTCX-2015-ZD2)

摘　　要：目的构建融合表达幽门螺杆菌(H.pylori)抗原Ure B与Hsp A的乳酸乳球菌(L.lactis)菌株,为H.pylori感染的疫苗和免疫诊断研究奠定基础。方法从前期构建的hsp A-ure B融合基因克隆菌株E.coli TB1/p MAL-c2x-hsp Aure B中提取质粒,用PCR方法从质粒中扩增hsp A-ure B基因,将扩增产物经限制性酶切后与表达载体p NZ8110连接,用于转化E.coli MC1061菌株,从转化子中提取重组质粒p NZ8110-hsp A-ure B,用电穿孔法将其转入L.lactis NZ3900菌株中,用乳链菌素诱导hsp A-ure B表达,应用PAGE分析表达产物。结果 hsp A-ure B基因的PCR扩增产物长度为2 085 bp,酶切、PCR和测序鉴定结果均显示L.lactis表达系统构建正确,PAGE分析未能确定融合基因表达条带。结论首次以L.lactis为宿主菌成功构建H.pylori 2种抗原的融合表达系统,研究发现对H.pylori口服疫苗和诊断技术发展具有重要意义。Objective To construct a Lactococcus lactis（ L. lactis） strain expressing Helicobacter pylori（ H. pylori） Hsp A and Ure B in fusion to lay foundation for the development of H. pylori vaccine and diagnostic techniques. Methods The fusion gene hsp A- ure B was obtained by PCR amplification from the plasmid isolated from a recombinant E. coli strain TB1 / p MAL- c2x-hsp A- ure B,and the PCR product ligated to plasmid p NZ8110 was used to transform E. coli MC1061,and then the recombinant plasmid p NZ8110- hsp A- ure B isolated from the transformants was electrotransformed into L. lactis NZ3900. The L. lactis transformants were induced to express the hsp A- ure B gene by using nisin. And the expression produce was analyzed by PAGE.Results The PCR product of hsp A- ure B gene was 2 085 bp in length. The L. lactis expression system of hsp A- ure B gene was correctly constructed as identified by restriction enzyme digestion analysis and gene sequencing. No strips were recognized as the expression product of the fusion gene in PAGE pattern. Conclusion The fusion expression system of two H. pylori antigens was successfully constructed with L. lactis as host for the first time,which is of significance for advance in vaccine and diagnosis methods for H. pylori infection.

关 键 词：乳酸乳球菌 热休克蛋白 尿素酶 幽门螺杆菌 融合表达 

分 类 号：R378.2[医药卫生—病原生物学]