鸭肠炎病毒SORF3基因的克隆及表达载体的构建

Cloning of Duck Enteritis Virus SORF3 and Construction of an Expression Vector

克隆鸭病毒性肠炎病毒(Duck Enteritis Virus,DEV)SORF3基因并进行原核表达,为研究SORF3基因功能奠定基础。以DEV基因组为模板,PCR扩增SOFR3基因克隆至表达载体pET32a,构建重组表达质粒pET32a-SORF3,将重组质粒转化至感受态细胞BL21中,IPTG诱导表达并进行SDS-PAGE及Werstern-blot鉴定。结果构建的原核表达质粒pET32aSORF3转化BL21后经IPTG诱导4h,可表达分子量约50kDa融合蛋白,Western blot显示重组蛋白可以被Anti-His标签抗体识别,表明成功构建了pET32a-SORF3原核表达质粒,并能在大肠杆菌中表达具有免疫反应性的SORF3,为SORF3基因的功能研究奠定基础。

To clone the SORF3 gene of the duck enteritis virus （DEV） and express in prokaryotic cells to study the functional prerequisites for SORF3. The full-lenth SORF3 gene was amplified using the DEV genome as a template. The recombinant plasmid pET32a-SORF3 was constructed by cloning SOFR3 gene into the expression vector pET32a. The recombinant plasmid was transformed into competent cell BL21, protein expression was induced with IPTG and identified by SDS-PAGE and Werstern-blot. Analysis. Results indicated that the fusion protein with molecular weight of about 50KD was expressed with IPTG for 4 h. Western blot analysis showed that the recombinant protein could be recognized by Anti-His tag antibody. This study successfully constructed the recombinant prokaryotic expression plasmid pET32a-SORF3 and it expressed the immunogenic SORF3 in E. coli, which lays a foundation for the functional study of SORF3 gene.