AsiaⅠ型口蹄疫病毒基因工程多肽疫苗的构建及其免疫原性分析  

The Recombinant Peptide Vaccines against Foot-andMouth Disease Virus Serotype AsiaⅠand Their Efficacy in Guinea Pigs

作　　者：韩姬[1] 王加龙[1] 焦晔[1] 刘明秋[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工程国家重点实验室,上海200433

出　　处：《复旦学报（自然科学版）》2008年第5期553-560,共8页Journal of Fudan University：Natural Science

基　　金：上海市国内科技合作资助项目(055458013)

摘　　要：为了构建AsiaⅠ型口蹄疫病毒多肽疫苗,用反转录PCR方法扩增VP1基因并测得该基因的核苷酸序列.将VP1(133AA^163AA)、VP2(1AA^33AA)抗原表位基因分别与β-半乳糖苷酶基因(简称LacZ’)3′末端相连构建表达质粒LacZ’-VP1和LacZ’-VP2.化学合成VP1(133AA^163AA)-VP2(1AA^33AA)-VP1(133AA^163AA)串联重复抗原表位DNA,然后分别连接在β-半乳糖苷酶基因和IgG重链恒定区基因的3′末端,构建两种重组表达质粒LacZ’-VP1(133AA^163AA)-VP2(1AA^33AA)-VP1(133AA^163AA)(命名为pT1)和IgG-VP1(133AA^163AA)-VP2(1AA^33AA)-VP1(133AA^163AA)(命名为pT2).通过Western blot、血清中和抗体效价测定、T细胞增殖反应及豚鼠抗病毒保护实验分析所构建疫苗的免疫原性及有效性.结果显示pT1、pT2表达的融合蛋白能够诱发豚鼠产生较高的中和抗体及刺激豚鼠T细胞增殖.pT1、pT2两种融合蛋白分别2次免疫豚鼠后,100%的豚鼠能抵抗100 ID50AsiaⅠ型口蹄疫病毒攻击.pT2融合蛋白1次免疫豚鼠后70%的豚鼠能抵抗100 ID50AsiaⅠ型口蹄疫病毒攻击.For constructing novel peptide vacdnes, RNA of FMDV serotype Asia Ⅰ AKT strain was isolated and reversetranscripted into cDNA. The nudeotide sequences of VP1 protein 133AA to 163AA and VP2 protein 1AA to 33AA were amplified and ligated to the 3＇ terminal of the β-galactosidase gene respectively to construct two different expression plasmids named LacZ＇-VPI and LacZ＇-VP2. The tandem sequence of VP1 （ 133AA- 163AA）-VP2 （ 1AA- 33AA）-VP1 （ 133AA- 163AA） was synthesized by chemical method and further ligated to β-galaetosidase gene to construct an expression plasmid named pT1. In the same way the tandem sequence was ligated to bovine IgG heaw chain constant region to construct another expression plasmid pT2. The efficacy of the vaccines was investigated by neutralization antibody assay, lympho-proliferation assay and viral challenge assay. The results showed that both pT1 and pT2 fusion proteins can significantly stimulate cellular and humoral response. The results of viral challenge showed that pT1 and pT2 could protect guinea pigs with 100% protection rate against 100 ID50 viral challenge with FMDV serotype Asia Ⅰby the twice vaccination method, but then only 70% by one time vaccination with pT2.

关 键 词：FMDV ASIA Ⅰ型 抗原表位 重组多肽疫苗 

分 类 号：Q78[生物学—分子生物学]