犬Serinc5对HIV-1复制能力影响的研究

发布时间：2018-08-08 19:38

【摘要】：丝氨酸整合因子5(Serinc5)是最新报道能够被携带到病毒粒子中的宿主限制性因子,具有抵抗人类免疫缺陷病毒I型(HIV-1)感染性的作用。与人Serinc5(hSerinc5)具有高度同源性的犬Serinc5存在两种形式,一种为全长基因组编码的犬Serinc5(cSerinc5),另一种为转录剪接体(cSerinc5X2),该剪接体在N端缺少1~37位氨基酸。为鉴定这两种犬Serinc5是否具有抑制HIV-1的复制能力,本研究从犬肾细胞系MDCK中提取犬总RNA,经RT-PCR分别扩增两种犬Serinc5的cDNA,并克隆到pcDNA3.1载体中,构建编码两种犬Serinc5的重组质粒。将这两种重组质粒分别与编码HIV-1野生型或Nef缺失型的质粒共转染293T细胞,收获所产生的HIV-1病毒粒子,通过ELISA方法检测到这两种犬Serinc5对HIV-1病毒粒子的产量无影响。进一步将病毒粒子分别感染TZM-bl细胞,通过荧光强度检测结果显示,与对照组相比,cSerinc5X2对HIV-1感染性无显著抑制作用,而携带cSerinc5的HIV-1感染性与携带cSerinc5X2的病毒和对照组相比降低约9倍(p0.01),表明cSerinc5的N端对于病毒感染活性的抑制具有关键作用。本研究为鉴定Serinc5抗病毒的功能区提供实验依据。
[Abstract]:Serine integration factor 5 (Serinc5) is a new host restriction factor which can be carried into virus particles. It can resist the infection of human immunodeficiency virus type I (HIV-1). There are two forms of canine Serinc5 with high homology with human Serinc5 (hSerinc5), one is dog Serinc5 (cSerinc5) encoding full-length genome, the other is transcriptional splice (cSerinc5X2). In order to determine whether the two kinds of canine Serinc5 have the ability to inhibit the replication of HIV-1, we extracted the total HIV-1 from the dog kidney cell line MDCK, amplified the cDNAs of two kinds of dog Serinc5 by RT-PCR, cloned them into pcDNA3.1 vector, and constructed the recombinant plasmid encoding two kinds of canine Serinc5. The two recombinant plasmids were cotransfected into 293T cells with HIV-1 wild-type or Nef deletion type plasmids, and the resulting HIV-1 virus particles were harvested. The two kinds of canine Serinc5 were detected by ELISA method to have no effect on the production of HIV-1 virus particles. The virus particles were further infected with TZM-bl cells respectively. The results of fluorescence intensity test showed that cSerinc5X2 had no significant inhibitory effect on HIV-1 infection compared with the control group. The infection of HIV-1 carrying cSerinc5 was about 9 times lower than that of the virus carrying cSerinc5X2 (p0.01), which indicated that the N-terminal of cSerinc5 played a key role in inhibiting the viral infection activity. This study provides experimental evidence for identification of Serinc5 antiviral functional regions.
【作者单位】：中国农科院哈尔滨兽医研究所-美国密歇根州立大学天然免疫联合实验室兽医生物技术国家重点实验室;中国农业科学院兰州兽医研究所;
【基金】：黑龙江省自然科学基金面上项目(H201602) 黑龙江省人力资源和社会保障厅留学回国人员择优资助项目(2015424)
【分类号】：S852.65

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