IBD分子佐剂靶向提呈亚单位疫苗重组载体的构建与表达  

The construction and expression of recombinant vectors of infectious bursal disease molecular adjuvant targeting the proposed subunit vaccine

作　　者：韩小英[1] 成大荣[1] 王永娟[2] 朱善元[2] 金鑫[1] 左伟勇[2] 洪伟鸣[2] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏畜牧兽医职业技术学院,江苏泰州225300

出　　处：《扬州大学学报（农业与生命科学版）》2013年第2期11-15,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏省农业科技支撑计划项目(BE2011365)

摘　　要：为制备安全、高效的传染性法氏囊病(IBD)分子佐剂靶向提呈亚单位疫苗,将去毒大肠埃希菌不耐热肠毒素(mLTA)、传染性法氏囊病病毒(IBDV)衣壳蛋白VP2、鸡淋巴细胞毒性T细胞相关抗原4(CTLA-4)胞外区的基因片段依次串联,构建原核表达质粒pET30a-mLTA-VP2-CTLA-4-MCS-IgV。重组质粒转化大肠杆菌BL21菌株后,通过限制性内切酶酶切鉴定以及序列分析,筛选到含有目的重组质粒的重组菌。对重组菌诱导培养后,通过SDS-PAGE证明其可以表达约93ku的融合蛋白,与预期大小(92.7ku)相当,且经过Western-blot鉴定融合蛋白具有良好的抗原性。该重组融合蛋白的成功表达为进一步研究IBD分子佐剂靶向提呈亚单位疫苗奠定基础。To develop a safe and efficient subunit vaccine of Infectious Bursal Disease, the study cloned three genes which were mLTA gene, VP2 gene of infectious bursal disease virus and chicken cytotoxic T lymphocyte-associated anti- gen-4 gene into pET vector and constructed the prokaryotic expression plasmid pET30a-mLTA-VP2-CTLA-4-MCS-IgV. After the transformation of the recombinant plasmid into BL21, positive clones were screened by restriction enzyme diges- tion identification and sequence analysis. SDS-PAGE assay proved that the positive clone can express 93 ku fusion protein after induction, which was accordance with our expectation （92.7 ku）, and after the identification of Western-blot fusion protein had good antigenicity. The successful expression of the fusion protein laid foundation for the further development of the molecular adjuvant and antigen targeting subunit vaccine of IBD.

关 键 词：传染性法氏囊病 分子佐剂 亚单位疫苗 重组载体 

分 类 号：S858.31[农业科学—临床兽医学]