肺炎链球菌自溶酶(LytA)蛋白B细胞表位的预测、筛选及确定  

Predication,selection and identification of B cell epitopes of LytA protein from Streptococcus pneumoniae

作　　者：麦璟莹[1,2] 李祥俊[3] 马长玲[1] 袁竹青[1] 

机构地区：[1]广州医科大学病原生物学与免疫学教研室,广东广州510182 [2]广州医科大学形态学实验中心,广东广州510182 [3]广州医科大学

出　　处：《热带医学杂志》2015年第3期313-318,共6页Journal of Tropical Medicine

基　　金：国家自然科学基金(81072491);广州市高校科技基金(10A173)

摘　　要：目的确定肺炎链球菌自溶酶(LytA)蛋白B细胞线性表位。方法通过生物信息学方法预测肺炎链球菌LytA蛋白分子的B细胞表位并人工合成相应肽段;经克隆、表达、纯化等步骤得到纯化的重组LytA(rLytA)蛋白用以免疫小鼠并进行Westen blot鉴定;取免疫组小鼠和对照组小鼠血清分别与人工合成的候选B细胞表位肽段进行间接ELISA检测,筛选出B细胞表位。结果利用多种方法分析LytA蛋白分子的B细胞表位,经整理得到11条候选肽段,命名为LB1到LB11,并进行了人工合成。利用分子生物学技术获得了rLytA蛋白并成功地免疫了小鼠,间接ELISA结果显示:肽段LB3、LB5、LB6、LB7、LB11与rLytA免疫小鼠的血清发生反应,其OD 450 nm读数比相应的阴性对照小鼠显著升高,差异有统计学意义(P<0.01)。结论确定了肺炎链球菌LytA蛋白分子5个B细胞线性表位的位置和序列。Objective To identify B cell leanerepitopes of Streptococcus pneumoniae LytA protein. Methods Bioinformatics methods were used to predict B cell epitopes of Streptococcus pneumoniae LytA protein,and the B cell epitope peptides were synthesized. Purified rLytA was used to immunize mice. Sera were collected from each of immunized mice and negative control mice and used to perform indirect ELISA with synthesized peptides. Results Several bioinformatics methods were used to predict B cell epitope of LytA and 11 peptides were chosen as candidate B cell epitopes, which named from LB1 to LB11,and the peptides were synthesized. By using molecular biology technologies, we obtained purified rLytA protein and successfully immunized mice. Indirect ELISA showed that peptide LB3, LB5, LB6, LB7 and LB11 reacted with immunized mice serum and the OD450 nm values were significant higher than those of the unimmunizedmice. Conclusion Five B cell leaner epitopes of Streptococcus pneumoniae LytA protein were identified.

关 键 词：肺炎链球菌 自溶酶 B细胞表位 

分 类 号：R378.12[医药卫生—病原生物学]