Abstract
Cytomegalovirus (CMV) promoter drives various gene expression and yields sufficient protein for further functional investigation. Receptor binding domain (RBD) on spike protein of the SARS_CoV2 is the most critical portal for virus infection. Thus native conformational RBD protein may facilitate biochemical identification of RBD and provide valuable support of drug and vaccine design for curing COVID-19. We attempted to express RBD under CMV promoter in vitro, but failed. RBD-specific mRNA cannot be detected in cell transfected with recombinant plasmids, in which CMV promoter governs the RBD transcription. Additionally, the pCMV-Tag2B-SARS_CoV2_RBD trans-inactivates CMV promoter transcription activity. Alternatively, we identified that both Chicken β-actin promoter and Vaccinia virus-specific medium/late (M/L) promoter (pSYN) can highly precede SARS_CoV2 RBD expression. Our findings provided evidence that SARS_CoV2 RBD gene can be driven by Chicken β-actin promoter or Vaccinia virus-specific medium/late promoter instead of CMV promoter, thus providing valuable information for RBD feature exploration.
Keywords: CMV promoter; RBD; SARS_CoV2; Transcription; Translation.
【저자키워드】 Transcription, RBD, translation., CMV promoter, SARS_CoV2, 【초록키워드】 COVID-19, Gene Expression, translation, Vaccine design, Transcription, in vitro, Spike protein, Receptor binding domain, Protein, mRNA, cytomegalovirus, CMV, vaccinia virus, virus infection, information, Critical, RBD protein, chicken, Evidence, CMV promoter, can not, portal, Support, β-actin, biochemical, binding domain, Express, vaccinia, plasmids, promoter, Cell, RBD gene, provided, facilitate, functional, the RBD, conformational, driven by, RBD expression, transfected with, 【제목키워드】 RBD gene, driven by,