传染性囊病病毒VP2蛋白诱导细胞凋亡的研究

Studies on Apoptosis Induced by VP2 of Infectious Bursal Disease Virus

在体外分别构建了 3种重组表达质粒 :含传染性囊病病毒GZ911株VP2基因的pGVP2、含HK4 6株VP2基因的pHVP2以及含HK4 6株 5’ -端序列 (包括非编码区、VP5和VP2基因 )的pHVP2 +5 .用不同的重组质粒转染原代鸡胚成纤维细胞 (CEF) ,在转染后 0、14、38h ,收集对照组和转染组细胞 ,用酶联免疫吸附试验检测CEF中降解DNA量 .结果 ,在 14和 38h ,各组细胞中测得的Dλ 值均为转染pHVP2 +5组最大 ,其次为转染pHVP2、pGVP2的组 ,并都大于其他对照组 ,表明 2个IBDV毒株的VP2蛋白在CEF中表达后都能诱导CEF发生凋亡 ,但不同毒株的VP2蛋白诱导CEF凋亡的能力不同 ;

Three recombinant plasmids,pGVP2 and pHVP2 containing VP2 gene of IBDV strain GZ911 and HK 46 respectively, and pHVP2+5 containing 5' terminal sequence including 5' non coding region,VP5 gene and VP2 gene, were cloned. These three plasmids were used to transfect chicken embryo fibroblasts(CEF). At 0,14,38 h post transfection, cells in different groups were collected and the amount of DNA fragments was measured with enzyme linked immunosorbent assay(ELISA). At 14 and 38 h posttransfection, cells transfected with pHVP2+5 produced the highest D λ , cells transfected with pHVP2 produced intermediate D λ , and pGVP2 produced the lowest D λ . All of which were higher than control groups. These results showed that VP2 of two IBDV strains was able to induce apoptosis when expressed in CEF, but the VP2 of different strains varied in the capacity to induce apoptosis. The apoptosis in CEF induced by VP2 was enhanced by VP5.