CAJ | 학술논문

目的探讨运用逆向疫苗学理论,筛选沙眼衣原体疫苗候选蛋白的可行性. 方法通过对沙眼衣原体测序基因组中890个基因进行生物信息学分析,预测编码表面暴露蛋白的基因,进一步利用实时定量PCR、蛋白质表达纯化、免疫动物以及特异性抗体效价的测定,验证候选蛋白的免疫原性. 结果经过生物信息学分析,结构预测及亚细胞定位,共得到126个符合要求的蛋白. 排除与人体可能存在同源的蛋白后,共得到73个候选蛋白. 利用实时定量PCR分析其中30个蛋白编码的基因,挑选其中表达量比较高的4个进行免疫效果的评价. 实验证实这4个基因编码的蛋白均可以产生保护性抗体. 结论本研究证实逆向疫苗学理论在沙眼衣原体疫苗的抗原筛选工作中是可行的,为下一步寻找有效的候选疫苗抗原打下了良好的基础.
목적 탐토운용역향역묘학이론,사선사안의원체역묘후선단백적가행성. 방법 통과대사안의원체측서기인조중890개기인진행생물신식학분석,예측편마표면폭로단백적기인,진일보이용실시정량PCR、단백질표체순화、면역동물이급특이성항체효개적측정,험증후선단백적면역원성. 결과 경과생물신식학분석,결구예측급아세포정위,공득도126개부합요구적단백. 배제여인체가능존재동원적단백후,공득도73개후선단백. 이용실시정량PCR분석기중30개단백편마적기인,도선기중표체량비교고적4개진행면역효과적평개. 실험증실저4개기인편마적단백균가이산생보호성항체. 결론 본연구증실역향역묘학이론재사안의원체역묘적항원사선공작중시가행적,위하일보심조유효적후선역묘항원타하료량호적기출.
Objective To investigate the feasibility of screening candidate proteins of Chlamydia pneumoniae vaccine based on the theory of reverse vaccine. Methods Bioinformation of 890 genes in the whole genome of Chlamydia pneumonia was analyzed to predict genes coding surface exposed proteins. The immunogenicity of candidate proteins was verified by real time quantitative PCR, proteins expression and purification, determination of antibody titers and specificity. Results Based on bioinformatics analysis, structure prediction and subcellular localization, 126 proteins were selected. After ruling out potential humanity homologous proteins, 73 candidate proteins were obtained. Among them, 30 hypothetical proteins were analyzed with real time quantitative PCR, the immune effects of 4 higher expression proteins were assessed with animal experiments, and the ultimate results showed that 4 proteins coded with high-expressed genes induced the production of protective antibodies. Conclusion The research confirms the feasibility of reverse vaccinology theory for screening Chlamydia vaccine candidate antigens and contributes to the further effort for acquiring effective candidate antigen.