摘要
目的构建表达幽门螺杆菌(H.pylori)中性粒细胞激活蛋白A(NapA)的重组乳酸乳球菌菌株,为研究NapA作为疫苗抗原和免疫调节剂的应用效果奠定基础。方法用PCR方法从H.pylori基因组DNA中扩增napA基因,将napA经Nae I和SphⅠ双酶切与表达载体pNZ8110连接,用电穿孔法转入乳酸乳球菌菌株NZ3900中,并通过SDS-PAGE和Western blots分析鉴定napA表达产物。结果扩增的nap A基因长度为435 bp,构建的重组乳酸乳球菌经nisin诱导可表达分子量约为19 kD的重组蛋白,表达量约占菌体总蛋白量的12.4%,该重组蛋白可被小鼠抗H.pylori血清识别。结论成功构建了表达NapA的重组乳酸乳球菌菌株,该菌株在H.pylori口服疫苗和黏膜免疫调节研究中具有应用潜力。
Objective To construct a expression system of Helicobacter pylori NapA gene in Lactococcus lactis, so as to lind ba- sis for developing Napa as H. pylori vaccine and immunoregulator. Methods The napA gene was obtained by PCR amplification from the genomic DNA of H. pylori, and ligated to the expression vector plasmid pNZ8110. The plasmid pNZSll0 -napA was electrotransformed into L. lactis NZ3900. The expression products of napA were analyzed by SDS - PAGE and Western blots. Results The length of amplified napA gene by PCR was 435 bp. The construction of recombinant Lactctococcus lactis expression, induced by nisin, showed that the recombinant protein molecular weight was about 19 kd, accounting for approximately 12.4% in the cell lysates, which could be recognized by anti -H. pylori mouse sera. Conclusion The recombinant strain of L. lactis capable of expression of napA has been successfully constructed, showing potential for approach to anti - H. pylori vaccine and mucosal immunoregulatory mechanism.
作者
彭晓燕
张荣光
段广才
孙楠
王琛
张凌寒
PENG Xiao -yan ZHANG Rong - guang DUAN Guang - cai SUN Nan WANG Chen ZHANG Ling -han(Department of Epidemiology and Statistics, College of Public Health, Zhengzhou University, Zhengzhou, Henan 450001, China)
出处
《中国卫生检验杂志》
CAS
2017年第10期1372-1374,共3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)
