本文關(guān)鍵詞： 核苷二磷酸激酶B 表達(dá) 新城疫病毒 復(fù)制與增殖　出處：《中國(guó)畜牧獸醫(yī)》2017年08期 　論文類型：期刊論文

【摘要】：為研究核苷二磷酸激酶B(nucleoside diphosphate kinase B,NDPK B)蛋白表達(dá)對(duì)新城疫病毒(Newcastle disease virus,NDV)F48E9株復(fù)制與增殖的影響,本試驗(yàn)將真核表達(dá)重組質(zhì)粒pEGFP-NDPK B轉(zhuǎn)染Vero細(xì)胞,經(jīng)G418壓力篩選出了陽(yáng)性細(xì)胞單克隆,并通過(guò)RT-PCR、Western blotting和倒置熒光顯微鏡觀察鑒定了NDPK B mRNA和蛋白質(zhì)的表達(dá),在細(xì)胞系基礎(chǔ)上,通過(guò)HA-HI、TCID50及實(shí)時(shí)熒光定量PCR檢測(cè)NDPK B蛋白對(duì)NDV F48E9株復(fù)制和增殖的影響。結(jié)果表明,試驗(yàn)成功地構(gòu)建了高表達(dá)NDPK B蛋白的Vero/NDPK B細(xì)胞系,并在此基礎(chǔ)上,通過(guò)檢測(cè)證明了NDPK B能抑制NDV F48E9株在Vero細(xì)胞中的復(fù)制與增殖,提示以NDPK B為基礎(chǔ)有可能設(shè)計(jì)開(kāi)發(fā)抗NDV的新藥物或抗病毒增效劑,對(duì)NDV進(jìn)行預(yù)防或治療。
[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.
【作者單位】： 河南省動(dòng)物疫病預(yù)防控制中心;吉林大學(xué)動(dòng)物醫(yī)學(xué)學(xué)院;
【基金】：河南省科技創(chuàng)新人才計(jì)劃項(xiàng)目(豫科人組[2011]1號(hào))
【分類號(hào)】：S852.65

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