检验医学与临床2024，Vol.21Issue(8)：1030-1034,5.DOI:10.3969/j.issn.1672-9455.2024.08.002
新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒的构建
Construction of MS2 phage virus-like particles with Omicron I1566V mutation site of SARS-CoV-2
摘要
Abstract
Objective To construct MS2 phage virus-like particles with Omicron I1566V mutation site of SARS-CoV-2.Methods The capsid protein(CP),mature protein(A protein)and Omicron I1566V mutant gene sequences of MS2 phage with 6×His tag were selected and synthesized,and inserted into the pACYCDu-et-1 plasmid to construct the recombinant vector.The recombinant protein was purified and characterized by transmission electron microscopy.Finally,the thermal stability and nuclease resistance of the virus-like parti-cles were detected by reverse transcription polymerase chain reaction(RT-PCR).Results The recombinant vector containing CP and A protein with 6×His tag and Omicron I1566V mutant gene sequence was success-fully constructed.The results were confirmed by restriction enzyme BamH I and Kpn I digestion and sequen-cing.After induction and purification,the virus-like particles with a diameter of 23-28 nm were observed by electron microscopy.The virus-like particles were digested by nuclease and stored stably at 37℃for more than 20 days.Conclusion In this study,the Omicron I1566V mutation site virus-like particles were success-fully constructed using the CP and A protein of bacteriophage MS2,which provided reliable quality assurance for the RT-PCR detection system of Omicron I1566V mutation site.关键词
新型冠状病毒变异株/Omicron I1566V突变位点/反转录聚合酶链反应/MS2噬菌体/病毒样颗粒Key words
SARS-CoV-2 variants/Omicron I1566V mutation/reverse transcriptase polymerase chain reaction/bacteriophage MS2/virus-like particles分类
医药卫生引用本文复制引用
杨静远,李永鑫,史茜,刘春燕,梁梦洁,张新..新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒的构建[J].检验医学与临床,2024,21(8):1030-1034,5.基金项目
国家临床重点专科建设项目[兵财社(2023)16号] （2023）
兵团临床医学研究中心建设项目[兵科发(2023)12号] （2023）
兵团重点领域科技攻关计划项目(2022AB030) （2022AB030）
新疆生产建设兵团医院院级科技计划项目(2022014). （2022014）
