本文選題：豬繁殖與呼吸綜合征病毒 + Nsp基因��； 參考：《華北農(nóng)學(xué)報(bào)》2017年01期

【摘要】：為了探索豬繁殖與呼吸綜合征病毒(PRRSV)強(qiáng)、弱毒株的Nsp9基因?qū)RRSV復(fù)制的影響,構(gòu)建含有PRRSV XH-GD強(qiáng)毒株、CH-1R弱毒株Nsp9全長基因的質(zhì)粒p IRes2-EGFP-Nsp9-XH-GD、p IRes2-EGFP-Nsp9-CH-1R,并將重組質(zhì)粒分別轉(zhuǎn)染到Marc-145細(xì)胞中,以MOI=1的劑量接種PRRSV XH-GD毒株,通過TCID50試驗(yàn)測定病毒的滴度,采用熒光定量PCR和Western Blot方法來測定病毒的N蛋白表達(dá)情況。結(jié)果表明,轉(zhuǎn)染弱毒株CH-1R Nsp9基因的Marc-145細(xì)胞中,PRRSV的N蛋白在mRNA水平、蛋白質(zhì)水平上的表達(dá)量均高于轉(zhuǎn)染強(qiáng)毒株XH-GD組,且病毒的滴度也顯著高于轉(zhuǎn)染強(qiáng)毒株XH-GD組。綜上,在Marc-145細(xì)胞中,弱毒株CH-1R Nsp9基因?qū)RRSV復(fù)制的促進(jìn)作用優(yōu)于強(qiáng)毒株XH-GD。
[Abstract]:In order to explore the effect of Nsp9 gene of porcine reproductive and respiratory syndrome virus (PRRSVV) and attenuated strain on PRRSV replication, the plasmid pIRes2-EGFP-Nsp9-XH-GDN pRes2-EGFP-Nsp9-CH-1R containing the full-length Nsp9 gene of PRRSV XH-GD virulent strain was constructed, and the recombinant plasmid was transfected into Marc-145 cells.The PRRSV XH-GD strain was inoculated with the dose of MOI=1, the titer of the virus was determined by TCID50 test, and the expression of N protein was detected by fluorescence quantitative PCR and Western Blot.The results showed that the expression of N protein in Marc-145 cells transfected with attenuated CH-1R Nsp9 gene was significantly higher than that in XH-GD group at the mRNA level, and the titer of the virus was significantly higher than that in the XH-GD group.In conclusion, in Marc-145 cells, the promotion of PRRSV replication by CH-1R Nsp9 gene of attenuated strain is superior to that of XH-GD strain.
【作者單位】： 華南農(nóng)業(yè)大學(xué)獸醫(yī)學(xué)院國家生豬種業(yè)工程技術(shù)研究中心廣東省動物源性人獸共患病預(yù)防與控制重點(diǎn)實(shí)驗(yàn)室;河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院;河南省農(nóng)業(yè)科學(xué)院農(nóng)業(yè)經(jīng)濟(jì)與信息研究所;河南牧業(yè)經(jīng)濟(jì)學(xué)院制藥工程學(xué)院;廣西欽州保稅港區(qū)出入境檢驗(yàn)檢疫局;
【基金】：國家自然科學(xué)基金項(xiàng)目(31272564) 公益性行業(yè)(農(nóng)業(yè))科研專項(xiàng)(201203039) 國家生豬現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系項(xiàng)目(CARS-36)
【分類號】：S852.651

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