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伪狂犬病毒(pseudorabies virus,PRV)是一种疱疹病毒,在自然界具有极广泛的宿主类群,是中国养猪业发展主要威胁之一。为了研究病毒的神经传导机制,实验采用Lipofectamine3 000转染试剂,将PP63质粒与伪狂犬病毒Fa株基因组DNA共转染BHK-21细胞,空斑筛选得到gI/gE/US9重组缺失病毒,并命名为SA215-T。采用PCR、基因测序、Western Blot、电镜检查和生长曲线测定等方法检测重组病毒PRV-SA215-T。研究结果显示,Western Blot未发现gE基因的表达,SA215-T株的电镜形态与野毒株无明显差异,SA215-T株与亲本毒株Fa株在细胞中的生长曲线差异不明显且均达到了较高的病毒滴度。
Pseudorabies virus (PRV), a herpes virus, has a very wide host population in nature and is one of the major threats to the development of China’s pig industry. In order to study the neurotransmitter mechanism of the virus, BHK-21 cells were co-transfected with PP63 plasmid and PRV Fa strain genomic DNA using Lipofectamine 3000 transfection reagent, and gI / gE / US9 recombinant deleted virus was screened by plaque screening. And named SA215-T. The recombinant virus PRV-SA215-T was detected by PCR, gene sequencing, Western Blot, electron microscopy and growth curve. The results showed that the expression of gE gene was not found in Western Blot, the electron microscopic morphology of SA215-T strain was not significantly different from that of wild-type strain, and the growth curve of SA215-T strain and Fa strain was not significantly different Higher virus titer.