Abstract
We have constructed a family of expression vectors containing a single transcription unit that is active in Escherichia coli, eukaryotic cells, and in coupled in vitro transcription-translation systems. These vectors use the Rous sarcoma virus-long terminal repeat (RSV-LTR) as the promoter/enhancer for eukaryotic cells. In vitro transcription is made possible by inclusion of a bacteriophage T7 promoter. This same promoter is actively transcribed in E. coli that produce T7 RNA polymerase. Other features of this transcription unit include a high-efficiency eukaryotic translation start codon, a phage f1 origin of DNA replication for site-directed mutagenesis and a three-frame stop codon that facilitates C-terminal deletion mutagenesis. We term this vector family, pEXPRESS.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Avian Sarcoma Viruses / genetics
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Base Sequence
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Cell-Free System
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Codon / genetics
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DNA Replication / genetics
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DNA-Directed DNA Polymerase / metabolism
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Escherichia coli / metabolism
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Genetic Vectors / genetics*
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HeLa Cells / metabolism
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Humans
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Molecular Sequence Data
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Mutagenesis, Site-Directed / genetics*
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Plasmids / genetics*
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Poly A / genetics
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Promoter Regions, Genetic / genetics
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Receptors, Thyroid Hormone / genetics
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Recombinant Proteins / biosynthesis*
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Repetitive Sequences, Nucleic Acid / genetics
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T-Phages / genetics
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Transcription, Genetic*
Substances
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Codon
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Receptors, Thyroid Hormone
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Recombinant Proteins
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Poly A
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DNA-Directed DNA Polymerase