猪圆环病毒2型ORF1和ORF3 T淋巴细胞抗原表位的克隆与原核表达

Cloning and Prokaryotic Expression of T Lymphocyte Epitopes on ORF1 and ORF3 of Porcine Circovirus Type 2

摘要: 根据Stevenson筛选出来的3个具有活性的T淋巴细胞抗原表位（T lymphocyte epitope，TCE）合成 tce 基因（180 bp），成功插入到pMD18-T Simple载体中,筛选获得重组质粒,命名为pMD-TCE.将 tce 酶切产物亚克隆到原核表达载体pET-32a（+）中,获得重组质粒,命名为pET-TCE.用IPTG进行诱导表达,收集菌液进行SDS-PAGE检测,结果显示合成基因在pET-32a（+）中获得了高效融合表达,其表达蛋白相对分子质量约为25 000, Western-blot分析结果表明,获得的融合蛋白可与猪圆环病毒2型(PCV2)阳性血清发生特异性反应,具有良好的免疫原性.

Abstract: According to three active T lymphocyte epitopes (TCE) which Stevenson screened, tce gene (180 bp) was synthesized and cloned successfully into pMD18-T Simple vector. tce gene was subcloned into prokaryotic expression vector pET-32a(+); the recombinant plasmid was named pET-TCE and induced by IPTG. The results of SDS-PAGE and Western-blot indicated that the tce gene was expressed at a high level, and the recombinant fusion protein was about 25 000. The recombinant fusion protein has a peculiar reaction to porcine circovirus 2 (PCV2) positive serum which has immunological activity.