中文摘要：

【目的】克隆和表达羊口疮病毒（ORFV）的B2L基因,进而纯化并分析其反应原性.【方法】根据GenBank已发表的ORFV（GQ328006）的B2L基因序列设计一对特异引物,以提取阳性临床样品的总DNA为模板,通过PCR方法获得ORFV的B2L基因,将该基因片段连接至原核表达载体pET-30a（＋）上,获得重组质粒pET-30a（＋）-B2L.将此重组质粒转化BL21（DE3）感受态细胞,挑取单克隆进行扩大培养,经IPTG诱导获得重组融合蛋白.用SDS-PAGE和Western-blotting对表达的目的蛋白进行分析.【结果】成功获得重组质粒pET-30a（＋）-B2L,读码框正确.获得了43ku的表达产物,与预期目的蛋白大小相符;纯化后的目的蛋白能与ORFV阳性血清发生特异反应.【结论】成功对ORFV-B2L蛋白进行了原核表达,且纯化后蛋白具有良好的反应原性.

英文摘要：

[Objective] To clone and express the B2L gene of ORFV, and then purify and analyze the re- actogenicity. [Method] A pair of primer was designed according to ORFV B2L gene reported in Gen-Bank （GQ328006）, taking the DNA of this virus extracted from the pathologic materials as template and the tar- get gene was got by PCR. After purification, the product was cloned into pET-30a（q-） vector to ob-tain the recombinant plasmid pET-30a（q-）-B2L. After identification, the recombinant plasmid was trans-formed in- to BL21（DE3） competent cells and the single colony was picked to amplify and culture. After induction with IPTG, the recombinant fusion protein was expressed. The target protein was identified by SDS-PAGE and Western blotting after purification. [Result]The results showed that the B2L gene was successfully cloned into pET-30a（q-）, a protein was confirmed at position about 43 kDa by SDS-PAGE and could specif- ically react with ORFV positive serum identified by western-blot. [Conclusion]This ex-periment realize the prokaryotic expression of protein B2L,and the purified protein had good reactogenicity.