猪圆环病毒3型7个ORFs不能诱导PK-15细胞凋亡

Seven open reading frames of porcine circovirus type 3 induced no apoptosis of PK-15 cells

【目的】为了探明猪圆环病毒3型的致病机制。【方法】用PCR技术扩增PCV3的ORF1-ORF7基因片段并克隆到pEGFP-C1载体,构建7个真核表达质粒后分别转染PK-15细胞,在转染后不同时间观察细胞中荧光表达情况,并检测Caspase-3活性变化。【结果】7个真核表达质粒转染PK-15细胞后均可观察到荧光,表明PCV3的ORF1-ORF7基因均可在PK-15细胞中获得表达,但在表达丰度方面存在差异。与单独的PK-15细胞和pEGFP-C1载体转染的PK-15细胞相比,7个真核表达质粒转染PK-15细胞后都未出现Caspase-3活性的显著(P>0.05)变化,表明均未引起PK-15细胞发生凋亡。【结论】PCV3的ORF1-ORF7基因均无诱导PK-15细胞发生凋亡的作用。

【Objective】 The aim of this study is to analyze whether the seven open reading frames (ORFs) of porcine circovirus type 3 (PCV3) are able to induce apoptosis of PK-15 cells or not.【Methods】 The ORF1-ORF7 genes of PCV3 was amplified by PCR method and cloned into pEGFP-C1 vector to construct 7 eukaryotic expression plasmids,respectively.These eukaryotic expression plasmids were transfected into PK-15 cells respectively.Their expressions in PK-15 cells were observed and the activities of Caspase-3 were detected at different times post transfection.【Results】 The results showed that the fluorescence signals were observed in PK-15 cells transfected with the whole of 7 eukaryotic expression plasmids,indicating that ORF1-ORF7 genes of PCV3 could be expressed in PK-15 cells with differences in expressions.There were no significant changes in Caspase-3 activities in PK-15 cells transfected with any of 7 eukaryotic expression plasmids compared with both PK-15 cells alone and PK-15 cells transfected with pEGFP-C1 vector,indicating that ORF1-ORF7 genes of PCV3 did not cause apoptosis of PK-15 cells.【Conclusion】 All the ORF1-ORF7 gene of PCV3 had not the ability to induce the apoptosis of PK-15 cells.