抗A5型口蹄疫病毒重组多肽疫苗的研究  

Study on the Recombinant Polypeptide Vaccine against Foot-and-mouth Disease Virus Type A5

作　　者：刘明秋[1] 张骏[1] 陈维灶[1] 严维耀[1] 郑兆鑫[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室,上海200433

出　　处：《复旦学报（自然科学版）》2005年第6期1037-1041,共5页Journal of Fudan University：Natural Science

基　　金：教育部重大资助项目(2001)

摘　　要：根据A型口蹄疫病毒(FMDV)的VP1基因序列及国际上公认的抗病毒中和表位,并结合对口蹄疫病毒的研究成果,设计了A型FMDV的重组多肽疫苗.分别选择VP1上21～40位和137～160位氨基酸对应的基因序列为表位基因,组成137～160-21～40-137～160的串连结构,并以大肠杆菌β-半乳糖苷酶为大分子载体,构建重组质粒pLM99,在大肠杆菌中表达得到高含量的融和蛋白.体外免疫原性检测表明,所表达的融和蛋白与标准A型阳性血清有特异性抗原/抗体反应,即说明该融和蛋白具有免疫反应性.免疫豚鼠的实验结果表明,融合蛋白能在豚鼠体内诱导中和抗体,并使70%的豚鼠能抵抗病毒的攻击.The peptide of amino acids 137 - 160 of VP1 protein of foot-and-mouth disease virus （FMDV） type A could be a major B cell epitope, and the peptide of amino acids 21 - 40 is an important T cell epitope. The DNA fragments coding for amino acids 137 - 160 and 21 - 40 of VP1 protein of type A FMDV were chemically synthesized and ligated into a tan- dem repeat 137- 160-21-40-137- 160. This tandem sequence was cloned to the 3＇ end of the β-galactosidase gene in plasmid pWR590 to form a fusion gene. Thus the prokaryotic expression plasmid pLM99 was constructed, pLM99 encodes a fusion protein of β-galactosidase and the tandem repeat epitopes. The immunogenicity and neutralizing antibody activities of the fusion protein were assayed. The Western blotting result showed that the fusion protein exhibited immunogenicity of FMDV. The fusion protein also elicited high antibody levels in guinea pigs in vivo and protected 70 percent guinea pigs against 100 LD50 viral challenge of FMDV type A.

关 键 词：A型口蹄疫病毒 抗原表位 多肽疫苗 

分 类 号：R329.7[医药卫生—人体解剖和组织胚胎学]