Translational efficiency of polycistronic mRNAs and their utilization to express heterologous genes in mammalian cells
- PMID: 3582359
- PMCID: PMC553375
- DOI: 10.1002/j.1460-2075.1987.tb04737.x
Translational efficiency of polycistronic mRNAs and their utilization to express heterologous genes in mammalian cells
Abstract
The translation of polycistronic mRNAs in mammalian cells was studied. Transcription units, constructed to contain one, two or three open reading frames (ORFs), were introduced stably into Chinese hamster ovary cells and transiently into COS monkey cells. The analysis of mRNA levels and protein synthesis in these cells demonstrated that the mRNAs transcribed were translated to generate multiple proteins. The efficiency of translation was reduced approximately 40- to 300-fold by the insertion of an upstream ORF. The results support a modified 'scanning' model for translation initiation which allows for translation initiation at internal AUG codons. High-level expression of human granulocyte-macrophage colony stimulating factor was achieved utilizing a vector that contains a polycistronic transcription unit encoding an amplifiable dihydrofolate reductase marker gene in its 3' end. Thus, polycistronic expression vectors can be exploited to obtain high-level expression of foreign genes in mammalian cells.
Similar articles
-
Improved vectors for stable expression of foreign genes in mammalian cells by use of the untranslated leader sequence from EMC virus.Nucleic Acids Res. 1991 Aug 25;19(16):4485-90. doi: 10.1093/nar/19.16.4485. Nucleic Acids Res. 1991. PMID: 1653417 Free PMC article.
-
Amplification and expression of human alpha-globin genes in Chinese hamster ovary cells.Mol Cell Biol. 1984 Aug;4(8):1469-75. doi: 10.1128/mcb.4.8.1469-1475.1984. Mol Cell Biol. 1984. PMID: 6493227 Free PMC article.
-
Initiation of translation at internal AUG codons in mammalian cells.Nature. 1984 May 3-9;309(5963):82-5. doi: 10.1038/309082a0. Nature. 1984. PMID: 6717585
-
Pushing the limits of the scanning mechanism for initiation of translation.Gene. 2002 Oct 16;299(1-2):1-34. doi: 10.1016/s0378-1119(02)01056-9. Gene. 2002. PMID: 12459250 Free PMC article. Review.
-
[Primary structure of mRNA and translation strategy of eukaryotes].Mol Biol (Mosk). 1987 Jul-Aug;21(4):888-914. Mol Biol (Mosk). 1987. PMID: 3309622 Review. Russian.
Cited by
-
Suppression of tumor formation in vivo by expression of the JE gene in malignant cells.Mol Cell Biol. 1991 Jun;11(6):3125-31. doi: 10.1128/mcb.11.6.3125-3131.1991. Mol Cell Biol. 1991. PMID: 2038321 Free PMC article.
-
Nuclear transcriptional activity of the tobacco plastid psbA promoter.Nucleic Acids Res. 1989 Jan 11;17(1):19-29. doi: 10.1093/nar/17.1.19. Nucleic Acids Res. 1989. PMID: 2563150 Free PMC article.
-
Male-enhanced antigen gene is phylogenetically conserved and expressed at late stages of spermatogenesis.Proc Natl Acad Sci U S A. 1989 Nov;86(21):8462-6. doi: 10.1073/pnas.86.21.8462. Proc Natl Acad Sci U S A. 1989. PMID: 2813404 Free PMC article.
-
Translation of a polycistronic mRNA in the presence of the cauliflower mosaic virus transactivator protein.EMBO J. 1991 Dec;10(12):3887-96. doi: 10.1002/j.1460-2075.1991.tb04958.x. EMBO J. 1991. PMID: 1935908 Free PMC article.
-
Interpreting cDNA sequences: some insights from studies on translation.Mamm Genome. 1996 Aug;7(8):563-74. doi: 10.1007/s003359900171. Mamm Genome. 1996. PMID: 8679005 Review.
References
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
