人致病性呼肠孤病毒μNS互作蛋白筛选与鉴定

Screening and identification of μNS interactive proteins of a human pathogenic strain of orthoreovirus

目的筛选呼肠孤病毒μNS蛋白的胞内未知互作蛋白并对其进行定性鉴定,为呼肠孤病毒药物的开发提供参考依据。方法将构建好的载有M3基因片段的质粒pEFHAM3及空质粒分别转染至293T细胞中,通过免疫沉淀方法得到M3基因片段编码的μNS蛋白及可能与之互作的宿主蛋白,对实验组及对照组蛋白分别进行蛋白质定性检测,分析差异蛋白。结果对所得蛋白样本进行蛋白质定性检测,得到丝氨酸/苏氨酸蛋白激酶、精胺合酶、角蛋白14、线粒体Rho GTPase 1、组蛋白H2B、40S核糖体蛋白S4、核糖体蛋白L26、肌动蛋白–2和未表征的蛋白质共9种差异蛋白。结论丝氨酸/苏氨酸蛋白激酶、精胺合酶、角蛋白14、线粒体Rho GTPase 1、组蛋白H2B、40S核糖体蛋白S4、核糖体蛋白L26、肌动蛋白–2和未表征的蛋白质中的1种或几种可能通过和μNS蛋白互作参与了呼肠孤病毒的复制。

Objective To screen and qualitatively identify unknown intracellular interactive proteins of Nelson Bay orthoreovirus μNS protein for providing a reference to the development of anti-reovirus drugs. Methods The plasmid pEFHAM3 and empty plasmids containing the M3 gene fragment were constructed and transfected into 293T cells, respectively. The μNS protein encoded by the M3 gene fragment and its interactive protein were obtained with immunoprecipitation method. The proteins of the two groups were detected qualitatively and differences in protein components were analyzed. Results Totally 9 differential proteins were qualitatively detected,including serine/threonine protein kinase,arginine synthase,keratin 14,mitochondrial Rho GTPase 1,histone H2B,40S ribosomal protein S4,ribose protein L26,actin-2,and an uncharacterized protein. Conclusion One or more of the nine differential proteins may be involved in the replication of Nelson Bay orthoreovirus inducing human respiratory infection.