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.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1016/s0168-1656(01)00388-1 | DOI Listing |
Front Immunol
December 2024
Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
Introduction: To analyze the molecular pathogenesis of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), a small animal model such as mice is needed: human angiotensin converting enzyme 2 (hACE2), the receptor of SARS-CoV-2, needs to be expressed in the respiratory tract of mice.
Methods: We conferred SARS-CoV-2 susceptibility in mice by using an adenoviral vector expressing hACE2 driven by an elongation factor 1α (EF1α) promoter with a leftward orientation.
Results: In this model, severe pneumonia like human COVID-19 was observed in SARS-CoV-2-infected mice, which was confirmed by dramatic infiltration of inflammatory cells in the lung with efficient viral replication.
Plant Dis
November 2024
Chinese Academy of Agricultural Sciences Grassland Research Institute, Hohhot, Inner Mongolia, China;
Environ Microbiol Rep
December 2024
Department of Natural Sciences and Mathematics, Pontificia Universidad Javeriana Cali, Cali, Colombia.
Vanilla is an economically important crop for low-lying humid tropical regions, but cultivated plants face serious phytosanitary problems. Fusarium wilt is a devastating disease affecting vanilla crops, caused by the fungal pathogens Fusarium oxysporum f. sp.
View Article and Find Full Text PDFPlant Dis
November 2024
Nanyang Institute of Technology, Nanyang, Henan, China;
Mycologia
November 2024
Department of Microbiology, Genetics & Immunology, Michigan State University, East Lansing, Michigan 48824.
Ectomycorrhizal fungi in the genus form hypogeous fruiting bodies called truffles. Many species are highly prized due to their edible and aromatic ascomata. Historically, there has been attention on cultivating and selling European truffle species, but there is growing interest in cultivating, wild-harvesting, and selling species of truffles endemic to North America.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!