新城疫病毒TL1株F、HN蛋白基因共表达对细胞融合的影响

目的分析新城疫病毒TL1株F、HN蛋白基因共表达对细胞融合的影响。方法在6孔细胞培养板上将4种重组表达质粒以pCI-M+pCI-NP+pCI-F和pCI-M+pCI-NP+pCI-F+pCI-HN两种组合分别共转染BHK-21细胞,转染后48h,光镜观察细胞融合现象。在25cm2细胞培养瓶中,将4种重组表达质粒以上述两种组合分别共转染BHK-21细胞,转染后48h,收集细胞上清,离心后,电镜观察有无病毒样颗粒。结果以pCI-M+pCI-NP+pCI-F+pCI-HN组合共转染BHK-21细胞,可以观察到明显的细胞融合现象,而以pCI-M+pCI-NP+pCI-F组合共转染的BHK-21细胞则未出现细胞融合现象。两种组合共转染均形成了新城疫病毒样颗粒,包含HN蛋白基因的4种重组质粒组合共转染形成的病毒样粒子更接近于真实的新城疫病毒粒子。结论HN蛋白基因具有促进细胞融合的功能,新城疫病毒样颗粒的成功表达为新城疫病毒致病机理和新型疫苗的研究奠定了基础。

Effect of Co-expressing F and HN Genes of Newcastle Disease Virus TL 1 Strain on Cell Fusion

Objective To investigate the effect of co-expressing F and HN genes of Newcastle disease virus(NDV)TL1 strain on cell fusion.Methods The BHK-21 cells on a 6-well culture plate were co-transfected with recombinant plasmids pCI-M + pCI-NP + pCI-F and pCI-M + pCI-NP + pCI-F + pCI-HN respectively and observed for cell fusion by light microscopy 48 h later.The BHK-21 cells in a 25 cm2 culture flask were co-transfected with recombinant plasmids pCI-M + pCI-NP + pCI-F and pCI-M + pCI-NP + pCI-F + pCI-HN respectively,and the culture supernatant was collected 48 h later and centrifuged,then the precipitate was resuspended and observed for presence or absence of virus-like particles(VLPs)by electron microscopy.Results Significant cell fusion was observed in BHK-21 cells transfected with recombinant plasmids pCI-M + pCI-NP + pCI-F + pCI-HN,while was not in those with pCI-M + pCI-NP + pCI-F.VLPs were formed in all the transfected cells,though those formed in the cells transfected with pCI-M + pCI-NP + pCI-F + pCI-HN were more similar to nature NDV particles.Conclusion HN gene promoted the cell fusion.The successful expression of NDV-like particles laid a foundation of study on pathogenic mechanism of NDV and development of novel NDV vaccine.