O型口蹄疫病毒VP1基因的克隆与表达

以O型口蹄疫病毒(FMDV)流行毒株为研究对象，通过RT-PCR扩增出VP1基因，克隆至表达载体pET-32a中，分析比较不同地区O型口蹄疫病毒VP1基因序列，为构建VP1重组基因工程苗奠定基础．经测序表明，目的基因VP1已以正确的阅读框架整合至表达质粒中，应用大肠杆菌BL21(DE3)为宿主菌，通过IPTG诱导方法，表达包含VP1基因产物的融合蛋白，经SDS-PAGE和Western—blotting分析，表明表达蛋白表达量高，反应原性良好．

Cloning and expression of foot-and-mouth disease virus type O VP1 gene

The subject of study was a foot-and-mouth disease virus(FMDV) type O prevalent strain. The VP1 gene of FMDV was amplified by RT-PCR and subsequently inserted into the expression vector pET-32a. The VP1 gene was sequenced and compared with other published FMDV type O strains isolated from different areas. The success in cloning of the VP1 gene laid down a basis for the construction of a recombinant genetically-engineered vaccine. Analysis of the sequence revealed that the reading frame of the VP1 gene insert was correct. The protein corresponding to the VP1 gene was expressed in E.coli BL21(DE3)after induction with IPTG. SDS-PAGE and western-blotting showed that the VP1 fusion protein was expressed efficiently and had good antigenicity.