肺炎链球菌蛋白PspC的制备与鉴定  

Preparation and identification of recombinant Streptococcus pneumoniae protein PspC

作　　者：谭亚军[1] 王丽婵 卫辰[1] 张华捷[1] 侯启明[1] 张庶民[1] 马霄[1] 

机构地区：[1]北京中国食品药品检定研究院百白破疫苗与毒素室/卫生部生物技术产品检定方法及其标准化重点实验室,100050

出　　处：《中国医药生物技术》2017年第5期397-401,共5页Chinese Medicinal Biotechnology

基　　金：国家高技术研究发展计划(863计划)(2012AA02A402)

摘　　要：目的对肺炎链球菌表面蛋白C(PspC)进行全基因合成、重组表达、纯化制备和鉴定。方法根据Gen Bank中Psp C的基因序列和氨基酸序列,通过密码子优化和全基因合成的方式获得目的蛋白基因,构建无标签、高效重组表达菌株,建立重组蛋白的纯化制备方法,并进行抗原鉴定及免疫原性检测分析。结果人工合成的Psp C基因序列经鉴定与预期一致,目的蛋白在大肠杆菌中获得了高效表达,表达量在30%以上,经两步纯化后纯度达到95%。重组蛋白能与肺炎链球菌阳性血清产生特异性抗原抗体反应,免疫小鼠后可诱导产生较高水平的蛋白特异性抗体。结论获得了重组肺炎链球菌蛋白Psp C,其具有与天然抗原相似的抗原性和免疫原性,为今后开展疫苗研制、载体蛋白应用等研究奠定基础。Objective To perform the gene synthesis, recombinant expression, preparation and identification of the Streptococcus pneumoniae protein Psp C. Methods The gene of Psp C was obtained by chemical synthesis after codons optimization according to the gene sequence and amino acid sequence in the Gen Bank. The label-free and efficient recombinant expression strain was constructed by molecular biological methods. The purification method of recombinant protein was established. And the protein＇s antigen identification and immunogenicity was tested. Results The synthetic gene sequence of Psp C was consistent with the database. The recombinant protein was highly expressed in E.coli. Expression of the target protein accounted for more than 30% of the total bacterial proteins. After purification by two-step chromatography, the protein purity reached 95%. The protein could react with human serum of clinically diagnosed pneumonia, indicating the protein had good antigenicity. Also high level of specific antibody could be induced after immunization with the protein in mice. Conclusion The recombinant Streptococcus pneumoniae protein Psp C is successfully prepared without label, which had similar antigenicity and immunogenicity as that of natural protein. This research provides foundation for future research on vaccine development and carrier protein application.

关 键 词：肺炎链球菌 重组蛋白质类 肺炎链球菌表面蛋白C 

分 类 号：R392-33[医药卫生—免疫学]