突变MARCH作用靶点在假病毒产率提升中的应用  

Improvement of the Yield of Pseudoviruses via Mutation of Lysine Residues Targeted by MARCH

作　　者：井汇源 段二珍 赵攀登[1] JING Hui-yuan;DUAN Er-zhen;ZHAO Pan-deng(Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;College of Biological Engineering,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南牧业经济学院,郑州450046 [2]河南工业大学生物工程学院,郑州450001

出　　处：《中国生物工程杂志》2023年第9期55-61,共7页China Biotechnology

基　　金：国家自然科学基金(32002265);河南省科技攻关(202102110095)。

摘　　要：抗病毒药物、中和抗体和预防性疫苗是应对新发、再发高致病性和高感染性病毒(如新冠病毒)最有效的策略。然而,涉及上述活病毒的实验操作必须在包含生物安全3级和4级设施的实验室中开展。为了便于评估上述抗病毒制品,研究者开发了基于人类免疫缺陷病毒(human immunodeficiency virus,HIV)和水疱性口炎病毒(vesicular stomatitis virus,VSV)骨架的假病毒包装系统。该系统同时包含靶标病毒的囊膜蛋白表达质粒,使包装好的假病毒具备与野生型病毒相似的受体吸附和膜融合功能。鉴于此,在特定的包装系统中,囊膜蛋白的包装过程对假病毒产率和感染性影响巨大。研究发现膜相关泛素连接酶家族蛋白(membrane-associated RING-CH,MARCH)能够降解病毒囊膜蛋白,下调假病毒产率和感染性。讨论了囊膜蛋白胞内域赖氨酸修饰对假病毒产率影响的研究进展,以期提高囊膜蛋白表达量和加工成熟效率,促进抗病毒药物研发、抗体筛选、疫苗创制和病毒受体鉴定。Therapeutic antiviral drugs,neutralizing antibody and preventive vaccines have been proven to be the most effective control measures towards emerging and reemerging viruses with high pathogenicity and infectivity like SARS-CoV-2.However,the experiments involving live viruses must be carried out in biosafety level 3(BSL-3)or BSL-4 facilities.To facilitate the evaluation of these antiviral products,the pseudovirus system has been developed based on the human immunodeficiency virus(HIV)/vesicular stomatitis virus(VSV)packaging system,as well as expression plasmids carrying envelope proteins to perform the function of attachment and fusion similar to the wild-type virus.Membrane-associated RING-CH(MARCH)E3 ubiquitin ligase proteins have been reported to downregulate the envelope proteins and significantly affect the yield and infection efficiency of pseudoviruses.This review summarizes the current progress in the effect of engineering MARCH-resistant envelop proteins by lysine modification on the production of pseudoviruses.Apparently,increased expression level from the surfaces of packaging cells and improved processing of the envelope protein in the packaging cells will greatly promote the development of antiviral drugs,antibody screening,vaccine research,and receptor recognition.

关 键 词：假病毒 包装系统 囊膜蛋白 赖氨酸修饰 

分 类 号：R373.2[医药卫生—病原生物学]