猪源新城疫病毒F蛋白裂解位点对BHK-21细胞特异性膜融合的影响

The effect of F protein schizolysis site gene of swine newcastle disease virus on specific membrane fusion of BHK-21 cell

目的研究猪源新城疫病毒F蛋白裂解位点区氨基酸序列对其特异性膜融合作用的影响。方法采用基因定点突变及基因重组的方法,测定突变猪源新城疫病毒JL01株F蛋白裂解位点区基因序列,并将阳性突变质粒与猪源新城疫病毒HN基因共转染BHK-21细胞,通过面积判断法和细胞计数法分析细胞融合效率。结果突变株RF4细胞融合效率为0.58±0.10,与野毒株的0.55±0.02接近。RF5融合效率较高,为0.69±0.05;RF3融合效率较低,为0.25±0.02。结论猪源新城疫病毒F蛋白裂解位点的改变对其特异性膜融合作用有一定影响。

Objective To investigate the effect of F protein schizolysis site gene of swine newcastle disease virus JL01 strain on specific membrane fusion. Methods Site-directed mutagenesis and gene recombinantion were used to get chimeric F protein.And then chimeric F protein and HN gene of swine Newcastle disease virus were co-transfected into BHK-21 cells.Giemsa staining and cell count method were used to analysis the fusion functions. Results The chimeric of RF4 is 0.58±0.10 had the same fusion functions as that 0.55±0.02,RF5 is 0.69±0.05,higher than that of JL01,RF3 is 0.25±0.02,lower than that of JL01. Conclusion The change of F protein schizolysis site gene of swine newcastle disease virus JL01 strain have some effects on specific membrane fusion.