【摘要】 目的构建淋球菌标准株外膜蛋白NspA基因重组子,在大肠杆菌BL21(DE3)中诱导表达外膜蛋白NspA。方法提取淋球菌标准株基因组DNA,PCR扩增其NspA基因,插入表达质粒载体pET-30c(+)中,构建pET-NspA重组子,转化表达宿主大肠杆菌BL21(DE3),异丙基-β-D-硫代半乳糖苷(IPTG)诱导重组蛋白表达,SDS-PAGE和WesternBlot检测表达蛋白。结果成功构建了淋球菌标准株的pET-NspA大肠杆菌表达重组子,经IPTG诱导表达后,该蛋白在大肠杆菌中高效表达。结论淋球菌外膜蛋白NspA在大肠杆菌中的成功表达,为研究该蛋白的免疫学性能、制备抗体和研制预防淋病的疫苗奠定了基础。更多还原

【Abstract】 Objective To construct neisseria surface protein (NspA) recombinants of Neisseria gonorrhoeae from a reference strain and express this protein in E.coli. Methods The fragments of NspA gene of Neisseria gonorrhoeae was amplified by PCR from the reference strain genomic DNA and cloned into expression vector pET-30c(+) to get the pET-NspA recombinants. The recombinants were verified with restrictive endonuclease digestion and sequence analysis. The verified recombinant was transformed into E.coli BL21 (DE3). After inducing with IPTG, the expressed NspA protein was analyzed by SDS-PAGE and Western Blot. Results The pET-NspA expression recombinants for the reference strain of Neisseria gonorrhoeae were successfully constructed and the induced recombinant NspA protein was observed. Conclusion The successful expression of the Neisseria gonorrhoeae NspA protein will be very helpful for the further research of its antigenicity and immunological activity, and for the construction of preventive vaccines on Neisseria gonorrhoeae infection.更多还原