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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。
The influenza virus nucleoprotein (NP), the major structural protein encoded by viral RNA segment 5, forms with the viral genomic RNA and polymerase (PB1, PB2, PA) a ribonucleoprotein (vRNP) complex that is the viral genome’s transcriptional and Copy of the basic functional units. In our laboratory, the yeast two-hybrid technique was used to screen the human nuclear cell cDNA library for the screening of the host protein U1 small nuclear ribonucleoprotein A (SNRPA), which interacts with the NP protein of influenza virus. In this study, direct interaction between NP and SNRPA was further confirmed by yeast backcross validation, co-immunoprecipitation (Co-IP) and GST pull down assay. Using siRNA interference gene expression, the replication titer of influenza virus decreased 2.7 times and 1.75 times respectively at 24 h and 48 h, indicating that SNRPA protein expression plays a positive regulatory role on influenza virus replication. This study enriched the protein regulatory network between the influenza virus protein and the host protein, which lays the foundation for further research on the mechanism of influenza virus replication.