Efficiency of different viral promoters in directing gene expression in mammalian cells: effect of 3'-untranslated sequences.
We compared (i) the enhancer/promoter (mCMV promoter) from the murine cytomegalovirus (CMV) major immediate early gene,(ii) the enhancer/promoter from human CMV major immediate early gene, containing a short promoter (h1CMV) or a long stretch of 5' untranslated region (UTR) from the gene promoter (h2CMV) and (iii) the simian virus 40 (SV40) enhancer/early region promoter (SV2) for their ability to direct foreign gene expression in transiently transfected mammalian cell lines. Two series of recombinant plasmids containing the different viral promoters fused to the cat reporter gene and 3'-UTR for processing of transcripts from either the SV40 early region or the rabbit Beta 1-globin-encoding gene (Glb) were also analyzed for their effect on transient gene expression. The mCMV was the most active in dihydrofolate reductase-deficient Chinese hamster ovary (CHOdhfr-) cells and BALB/3T3 clone A31 mouse embryo cells. The h2CMV was more active than the other promoters in Bowes human melanoma cells and in Vero African green monkey kidney cells. In human hepatoma (Hep G2) cells, similar levels of CAT synthesis were observed with the h2CMV- and the mCMV-based vectors. In Hep G2 and Bowes cells, 3'-UTR from the SV40 early region resulted in consistently higher levels of cat expression, as compared to the rabbit beta 1-Glb gene, while the converse was true in BALB/3T3 clone A31 and Vero cells. SV40 early region and rabbit beta1-Glb gene 3'-UTR resulted in similar cat expression in CHOdhfr- cells.
Rotondaro
L
L
Department of Biotechnology, Menarini Ricerche Sud, Pomezia (Roma), Italy.
Mele
A
A
Rovera
G
G
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D002999:Clone Cells; D006224:Cricetinae; D003587:Cytomegalovirus / Q000235:genetics*; D017931:DNA Primers; D004742:Enhancer Elements, Genetic; D015967:Gene Expression Regulation, Viral; D017781:Genes, Immediate-Early; D005822:Genetic Vectors; D006801:Humans; D008322:Mammals; D008545:Melanoma; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured; D014709:Vero Cells
D017931:DNA Primers; D011994:Recombinant Proteins