摘要
目的:对以轮状病毒(RV)重组VP6蛋白为载体插入Ⅲ型脊髓灰质炎病毒(PV3)VP1蛋白上1个抗原表位(VP1的91~102、254和168位氨基酸残基)所构建的嵌合蛋白6F/PV3N1进行体外免疫学研究。方法:利用分子克隆和基因重组技术将PV3抗原表位插入RV载体蛋白,构建重组抗原表位嵌合蛋白表达质粒,转染大肠杆菌后表达重组蛋白,经SDS-PAGE确认表达产物,再通过Western印迹分析嵌合蛋白的抗原反应性。结果:用载体蛋白VP6F、轮状病毒Wa病毒株免疫的豚鼠血清抗体分别都能与VP6F和6F/PV3N1产生特异性结合;PV3免疫的豚鼠血清抗体只能与6F/PV3N1产生特异性结合,不能与VP6F产生特异性结合;PV1免疫的豚鼠血清抗体则不能与6F/PV3N1和VP6F产生特异性结合。结论:PV1、PV3之间不存在交叉反应现象,以RV VP6为载体构建的嵌合蛋白6F/PV3N1具有较好的免疫原性,为研发RV/PV3嵌合疫苗提供了基础。
Objective: To study immunological activity in vitro of a chimeric protein constructed by inserting one recombinant epitope of VP6 of poliovirus typeⅢ(PV3)(amino acids 91~102, 254, and 168 of VP1) into the VP6 protein of recombinant poliovirus(RV) as a vector. Methods: PV3 epitope was inserted into the RV carrier protein by molecular cloning and gene recombination technology to construct a chimeric protein expression plasmid of the recombinant antigen epitope. After it was transfected into E.coli, it was expressed and followed by SDSPAGE test. Synthetic protein antigen reactivity was tested by Western blot. Results: The guinea pig serum antibodies immunized with the carrier protein VP6 F and rotavirus Wa strain can specifically bind to VP6 F and 6 F/PV3 N1,respectively. PV3-immune guinea pig serum antibodies can only specifically bind to 6 F/PV3 N1, but not to VP6 F.Specific binding occurs. PV1 immunized guinea pig serum antibodies do not specifically bind 6 F/PV3 N1 and VP6 F.Conclusion: There is no cross-reaction between PV1 and PV3. The chimeric protein 6 F/PV3 N1 constructed using RV VP6 as a vector has better immunogenicity. This study provides a basis for the development of RV/PV3 chimeric vaccines.
作者
杨筱舟
王晶
夏文跃
潘小霞
赵冰心
滕玉梅
文喻玲
陈元鼎
YANG Xiao-Zhou;WANG Jing;XIA Wen-Yue;PAN Xiao-Xia;ZHAO Bing-Xin;TENG Yu-Mei;WEN Yu-Ling;CHEN Yuan-Ding(Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Diseases, Kunming 650118;Key Laboratory of ChemistlT in Ethnic Medicinal Resources, State Ethnic Affairs Commission and Ministry of Education, Yunnan Minzu University, Kunming 650500 China)
出处
《生物技术通讯》
CAS
2018年第3期362-366,共5页
Letters in Biotechnology