猪圆环病毒Ⅱ型修饰Cap蛋白基因的原核表达及纯化

The prokaryotic expression and purification for the modified capsid gene of porcine circovirus 2

目的:本研究基于原核表达系统表达猪圆环病毒Ⅱ型(PCV-2)Cap蛋白。方法:PCV-2ORF2基因编码的Cap蛋白含有能产生免疫反应的多个抗原表位,本研究针对Cap蛋白基因设计引物,去掉核定位信号肽的核苷酸序列克隆到原核表达载体pET-32a(+)中,筛选重组菌并进行原核表达、鉴定及纯化。结果:纯化后的修饰Cap蛋白是能够与猪圆环病毒Ⅱ型的阳性血清发生特异性反应。结论:表达的重组修饰Cap蛋白可以作为标准抗原蛋白用于PCV-2的检测。

Objective： The study expressed the recombinant protein His-ORF2 （Cap） and subsequently purified it. Methods： The nucleic localization signal sequence was cut and the left nucleotide sequence of ORF2 of PCV-2 was cloned into the expression vector pET-32a（＋）.Results： The recombinant Cap protein can react with the PCV-2 positive serum in the western blot assay. Conclusion： The recombinant Cap protein can be used for the detection of PCV-2 infection.