新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒的构建OACSTPCD

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.