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目的:利用蛋白质相互作用的技术筛选与pUL23相互作用宿主蛋白,为研究pUL23蛋白对人巨细胞病毒繁殖的影响提供线索。方法:通过酵母双杂交系统从人胚肾cDNA文库筛选与pUL23相互作用的宿主蛋白;通过GST-pu ll-down技术研究二者体外物理性相互作用;免疫共沉淀技术进一步研究二者在胞内相互作用。结果:Pu ll-down技术、免疫共沉淀技术确定了宿主蛋白IGFBP4与pUL23具有相互作用。结论:上述结果为研究pUL23蛋白调节病毒自身繁殖功能提供重要依据。
OBJECTIVE: To screen the host proteins interacting with pUL23 by using the protein-protein interaction technology and provide clues for studying the influence of pUL23 protein on the human cytomegalovirus. METHODS: Host proteins interacting with pUL23 were screened from human embryonic kidney cDNA library by yeast two-hybrid system. The in vitro physical interaction between GFP and pUL23 was studied by GST-pu ll-down technique. The co-immunoprecipitation assay was used to further study the intracellular interaction. RESULTS: The Pu ll-down technique and co-immunoprecipitation technique identified the interaction of the host protein IGFBP4 with pUL23. Conclusion: The above results provide an important basis for the study of pUL23 protein regulation of the virus’s own reproductive function.