The human elongation factor 1 alpha (EF-1 alpha) first intron highly enhances expression of foreign genes from the murine cytomegalovirus promoter.
In order to develop a highly efficient mammalian expression vector, we constructed a vector by the combination of the murine cytomegalovirus (MCMV) immediate early (IE) promoter and the human elongation factor one alpha (EF-1 alpha) first intron. The MCMV IE promoter was several fold stronger than the human cytomegalovirus (HCMV) immediate early (IE) promoter and the human elongation factor one alpha (EF-1 alpha) promoter in various mammalian cell lines such as NIH3T3, Neuro-2a, 293T or HT1080 and was only slightly weaker than the HCMV or the EF-1 alpha promoter in HeLa and CHO cell lines. We inserted the first intron of the human EF-1 alpha gene behind the MCMV and the HCMV promoter to enhance the gene expression through increasing RNA transcription and/or RNA stability. The insertion of the human EF-1 alpha first intron markedly enhanced the level of gene expression in many cell lines and the resultant MCMV promoter with the human EF-1 alpha first intron (MCMV/EF-I) was higher than the HCMV promoter from 4.3 to 65.5-fold in various cell lines. Also, the MCMV/EF-I promoter induced higher level of erythropoietin expression than the HCMV promoter in transiently transfected CHO cells.
Kim
Seon-Young
SY
Laboratory of Cell Biology, Korea Research Institute of Bioscience and Biotechnology, 52 Eoundong, Yusong, Taejon 305-333, South Korea.
Lee
Jae-Ho
JH
Shin
Hyun-Seock
HS
Kang
Hye-Ja
HJ
Kim
Yeon-Soo
YS
D016475:3T3 Cells; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003587:Cytomegalovirus / Q000235:genetics*; D015870:Gene Expression; D017781:Genes, Immediate-Early / Q000235:genetics; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D051379:Mice; D020648:Peptide Elongation Factor 1 / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014162:Transfection
D020648:Peptide Elongation Factor 1