核苷二磷酸激酶B稳定表达细胞系的建立及其对新城疫病毒F48E9株复制的影响

发布时间：2018-02-24 03:07

本文关键词： 核苷二磷酸激酶B 表达新城疫病毒复制与增殖　出处：《中国畜牧兽医》2017年08期 　论文类型：期刊论文

【摘要】：为研究核苷二磷酸激酶B(nucleoside diphosphate kinase B,NDPK B)蛋白表达对新城疫病毒(Newcastle disease virus,NDV)F48E9株复制与增殖的影响,本试验将真核表达重组质粒pEGFP-NDPK B转染Vero细胞,经G418压力筛选出了阳性细胞单克隆,并通过RT-PCR、Western blotting和倒置荧光显微镜观察鉴定了NDPK B mRNA和蛋白质的表达,在细胞系基础上,通过HA-HI、TCID50及实时荧光定量PCR检测NDPK B蛋白对NDV F48E9株复制和增殖的影响。结果表明,试验成功地构建了高表达NDPK B蛋白的Vero/NDPK B细胞系,并在此基础上,通过检测证明了NDPK B能抑制NDV F48E9株在Vero细胞中的复制与增殖,提示以NDPK B为基础有可能设计开发抗NDV的新药物或抗病毒增效剂,对NDV进行预防或治疗。
[Abstract]:In order to study the effect of nucleoside diphosphate kinase (NDPK B) protein expression on replication and proliferation of Newcastle disease virus (NDV) F48E9 strain, the eukaryotic expression recombinant plasmid pEGFP-NDPK B was transfected into Vero cells. The expression of NDPK B mRNA and protein was observed by RT-PCR blotting and inverted fluorescence microscope. The effect of NDPK B protein on the replication and proliferation of NDV F48E9 strain was detected by HA-HI TCID50 and real-time fluorescence quantitative PCR. Vero/NDPK B cell lines with high expression of NDPK B protein were successfully constructed, and the results showed that NDPK B could inhibit the replication and proliferation of NDV F48E9 in Vero cells. The results suggest that it is possible to design and develop new drugs or antiviral synergists against NDPK B to prevent or treat NDV.
【作者单位】：河南省动物疫病预防控制中心;吉林大学动物医学学院;
【基金】：河南省科技创新人才计划项目(豫科人组[2011]1号)
【分类号】：S852.65

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