猪圆环病毒2型去核定位信号ORF2基因的原核表达与初步应用

Prokaryotic expression of NLS deleted ORF2 capsid protein of porcine circovirus type 2

根据PCV2-871病毒株的已知序列设计一对引物,PCR扩增其去核定位信号的Cap基因(ORF2),通过BamHⅠ/HindⅢ双酶切位点将其克隆到表达载体pQE-32中,转化大肠杆菌M15。经IPTG诱导表达、SDS-PAGE鉴定,表明去核定位信号的Cap蛋白可以大量表达。Western blot、ELISA鉴定蛋白活性,继而,以重组蛋白为抗原包被ELISA板,对28份临床样品进行检测。结果表明,该蛋白具有良好的抗原活性,可以作为抗原应用于临床样品的检测。

The NLS deleted ORF2 gene was amplified from PCV2-871 strain by PCR and cloned into pQE-32 vector. The recombinant plasmid pQE-PCV581 was transformed into E.coli M15 and protein expression was induced by IPTG. The expressed protein was show to react with PCV2 positive swine serum in ELISA and Western blot, and was successfully applied in ELISA to detect the clinic sera samples.