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.
出处
《扬州大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
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)
关键词
传染性法氏囊病
分子佐剂
亚单位疫苗
重组载体
infectious bursal disease
molecular adjuvant
subunit vaccines
recombinant vector
