鸡传染性贫血病毒VP2基因在真核细胞中的表达以及对宿主细胞增殖的影响

鸡传染性贫血是危害养禽业的重要疫病之一,其病原体是鸡传染性贫血病毒(CIAV)。该病原广泛存在于世界各地,是危害养禽业的主要病原。VP2是该病毒的重要非结构蛋白,其对宿主细胞的确切作用还不十分清楚。本文通过PCR从病毒DNA中扩增出VP2基因并将VP2基因亚克隆到真核表达载体pcDNA4.0。将重组质粒转染至BHK细胞,经Western-blotting证实,转染的真核细胞能表达VP2蛋白并具有良好的抗原性。pcDNA4.0-VP2转染293T或BHK细胞后,与对照组(空载体转染细胞以及未转染的细胞)相比,VP2超表达对这两种细胞的增殖没有影响。该结果表明鸡传染性贫血病毒的VP2基因不影响宿主细胞的增殖。

Expression of chicken infectious anemia virus (CIAV) VP2 gene in eukaryotic cells and its role in cell proliferation

Chicken infectious anemia is one of the most important diseases in poultry industry.The causative agent,chicken infectious anemia virus(CIAV),spread over the globe.The role of VP2,a non-structural protein of this virus,in the interaction between the virus and host cells was not clear. In this study,VP2 gene was cloned from the viral DNA by PCR,and then sub-cloned into the eukaryotic expression vector pcDNA4.0,which was examined by the restricted enzymatic analysis.The pcDNA4.0-VP2 plasmid was transfected into BHK or 293T cells for the expression of VP2.The expression of VP2 by the transfected cells was determined by Western-blot.To probe into the role of VP2 in cell proliferation,293T and BHK cells were transfected with pcDNA4.0-VP2 and examined by cell counting after Trypan blue staining.The result showed that over-expression of VP2 in 293T or BHK cells did not enhance the cell proliferation as compared to the controls,which indicated the uninvolvement of the viral VP2 in cell proliferation and divisions.