流感病毒NP蛋白与宿主蛋白SNRPA的相互作用研究

发布时间：2018-05-08 13:39

本文选题：酵母双杂交 + Co-IP　；参考：《中国预防兽医学报》2017年03期

【摘要】：流感病毒核蛋白(NP)是由病毒RNA节段5编码的主要结构蛋白,与病毒基因组RNA及聚合酶(PB1、PB2、PA)构成核糖核蛋白(vRNP)复合体,是病毒基因组的转录和复制的基本功能单位。本实验室采用酵母双杂交技术从人细胞系cDNA文库中筛选与流感病毒NP蛋白相互作用的宿主蛋白U1 small nuclear ribonucleoprotein A(SNRPA)。本研究通过酵母回交验证、免疫共沉淀(Co-IP)和GST pull down试验进一步证实NP与SNRPA之间存在直接相互作用。利用siRNA干扰基因的表达后,流感病毒的复制滴度在24 h和48 h分别下降2.7倍和1.75倍,表明SNRPA蛋白表达对流感病毒的复制发挥正调控作用。本研究丰富了流感病毒蛋白与宿主蛋白之间的蛋白质调控网络,为进一步研究流感病毒复制调控的机制奠定了基础。
[Abstract]:Influenza virus nucleoprotein (RNA) is a major structural protein encoded by RNA segment 5, which is composed of ribonucleoprotein (RNP) complex with viral genomic RNA and polymerase PB1 / PB2 (PAA). It is the basic functional unit for transcription and replication of viral genome. Yeast two-hybrid technique was used to screen the host protein U1 small nuclear ribonucleoprotein cDNA from human cell line cDNA library which interacted with NP protein of influenza virus. In this study, direct interaction between NP and SNRPA was further confirmed by yeast backcross test, immunoprecipitation Co-IP) and GST pull down test. The replication titer of influenza virus decreased 2.7 times and 1.75 times respectively at 24 h and 48 h after siRNA interference gene expression, indicating that the expression of SNRPA protein plays a positive role in influenza virus replication. This study enriches the protein regulatory network between influenza virus protein and host protein and lays a foundation for further study on the mechanism of influenza virus replication.
【作者单位】：中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部动物流感重点开放实验室;甘肃农业大学动物医学院;
【基金】：国家自然科学基金(31402206、31672582)
【分类号】：S852.65

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