本文關(guān)鍵詞： 豬流行性腹瀉病毒 非結(jié)構(gòu)蛋白 Ⅰ型干擾素 天然免疫應(yīng)答　出處：《畜牧獸醫(yī)學(xué)報(bào)》2017年03期 　論文類型：期刊論文

【摘要】：為研究豬流行性腹瀉病毒(PEDV)非結(jié)構(gòu)蛋白7(nonstructural protein 7,Nsp7)的亞細(xì)胞定位和對細(xì)胞Ⅰ型干擾素(IFN)應(yīng)答的影響,本試驗(yàn)利用生物信息學(xué)預(yù)測Nsp7基因序列,克隆了PEDV中國流行毒株Nsp7基因并構(gòu)建真核表達(dá)載體pCAGGS-Nsp7;采用Western blot和間接免疫熒光試驗(yàn)檢測Nsp7在細(xì)胞中的表達(dá)和分布;通過報(bào)告基因法、ELISA以及病毒復(fù)制抑制試驗(yàn)評(píng)估Nsp7對Ⅰ型IFN應(yīng)答的影響�；蚩寺『托蛄蟹治鼋Y(jié)果顯示PEDV Nsp7基因大小為249bp,在不同PEDV毒株間高度保守;Western blot結(jié)果顯示,Nsp7能夠在轉(zhuǎn)染細(xì)胞中高效表達(dá);間接免疫熒光試驗(yàn)顯示Nsp7主要定位在細(xì)胞質(zhì),僅少量分布在細(xì)胞核;雙熒光報(bào)告基因試驗(yàn)表明Nsp7能顯著抑制IFN-β啟動(dòng)子活性,ELISA結(jié)果顯示Nsp7能顯著抑制IFN-β蛋白的表達(dá),同時(shí)水皰性口炎病毒復(fù)制抑制試驗(yàn)顯示Nsp7明顯抑制poly(I:C)介導(dǎo)的Ⅰ型IFN的抗病毒作用。結(jié)果提示,Nsp7作為PEDV的保守蛋白,具有拮抗Ⅰ型IFN應(yīng)答的功能,為探討和揭示PEDV逃逸宿主天然免疫應(yīng)答的機(jī)制及指導(dǎo)臨床疫苗使用奠定基礎(chǔ)。
[Abstract]:For the study of porcine epidemic diarrhea virus (PEDV) nonstructural protein 7 (nonstructural protein 7, Nsp7) on the cell and subcellular localization of type I interferon (IFN) response of the test using bioinformatics to predict Nsp7 gene sequence of PEDV was cloned China strains Nsp7 gene and construct the eukaryotic expression vector pCAGGS-Nsp7; the expression and distribution of Western by blot and indirect immunofluorescence assay for the detection of Nsp7 in the cell; by reporter gene method, and the effect of ELISA virus replication inhibition test evaluation of Nsp7 on type I IFN responses. PEDV analysis showed that Nsp7 gene is 249bp gene cloning and sequence is highly conserved in different PEDV strains; Western blot the results showed that Nsp7 could be efficiently expressed in transfected cells; indirect immunofluorescence assay revealed that Nsp7 mainly localized in the cytoplasm, only a small amount of the distribution in the nucleus; luciferase test Show that Nsp7 can significantly inhibit IFN- beta promoter activity, ELISA results showed that the expression of Nsp7 can significantly inhibit IFN- protein, and vesicular stomatitis virus replication inhibition test showed that Nsp7 significantly inhibited poly (I:C) type antiviral effect mediated by IFN. The results suggest that Nsp7 as a conserved protein PEDV, antagonistic type the IFN response function, to explore and reveal the PEDV escape from the host innate immune response mechanism and clinical use of vaccines to lay the foundation.

【作者單位】： 河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院;
【基金】：河南省高校創(chuàng)新團(tuán)隊(duì)支持計(jì)劃(14IRTSTHN015) 河南省高等學(xué)校重點(diǎn)科研項(xiàng)目(16A230002)
【分類號(hào)】：S852.651

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 陳靜;楊盼盼;雷喜梅;王曉夢;高冬生;常洪濤;楊霞;陳陸;趙軍;王川慶;;PEDV中國流行株和疫苗株S蛋白免疫反應(yīng)性差異分析[J];中國獸醫(yī)學(xué)報(bào);2015年10期

2 劉艷成;杜雅楠;吳衛(wèi)杰;王雪飛;蘆婷;劉丹丹;;內(nèi)蒙古地區(qū)豬腹瀉相關(guān)病毒的檢測與分析[J];畜牧獸醫(yī)學(xué)報(bào);2015年09期

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 何余;;集束化護(hù)理在新生兒輸注高危藥物管理中的效果觀察[J];長江大學(xué)學(xué)報(bào)(自科版);2017年12期

2 yば踴，

本文編號(hào)：1532091