FEMS microbiology letters, 2002-11, Vol.216 (2), p.179-183
2002
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Details
- Autor(en) / Beteiligte
- Titel
- Overexpression in Escherichia coli of the AT-rich trpA and trpB genes from the hyperthermophilic archaeon Pyrococcus furiosus
- Ist Teil von
- FEMS microbiology letters, 2002-11, Vol.216 (2), p.179-183
- Ort / Verlag
- Oxford, UK: Blackwell Publishing Ltd
- Erscheinungsjahr
- 2002
- Quelle
- Wiley Online Library - AutoHoldings Journals
- Beschreibungen/Notizen
- Abstract Expression of AT-rich genes from microorganisms such as archaea is often inefficient in Escherichia coli. The trpA and trpB genes encoding the tryptophan synthase subunits were cloned from the hyperthermophilic archaeon Pyrococcus furiosus. No apparent difference in codon bias was found between the genes. However, using a conventional cloning vector having the lac promoter, the trpB gene was expressed poorly in E. coli, whereas the trpA gene was overexpressed. The expression of the trpB gene was remarkably enhanced (>12-fold) by the introduction of an overlapping leader open reading frame. The expression of the trpA gene was also improved (∼1.5-fold). This approach may be useful for overexpressing various kinds of AT-rich genes.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0378-1097eISSN: 1574-6968DOI: 10.1111/j.1574-6968.2002.tb11433.xTitel-ID: cdi_proquest_miscellaneous_18551818
- Format
- –
- Schlagworte
- Archaea, Archaeal Proteins - biosynthesis, Archaeal Proteins - genetics, Archaeon, AT Rich Sequence - genetics, AT‐rich gene, Bacteria, Bacteriology, Base Sequence, Biological and medical sciences, Cloning, Cloning vectors, Cloning, Molecular, Codon bias, Crystals, E coli, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Escherichia coli - genetics, Fundamental and applied biological sciences. Psychology, Gene Expression, Genes, Genetic Vectors - genetics, Genetics, Hyperthermophile, Microbiology, Microorganisms, Molecular Sequence Data, Mutagenesis, NADH, NADPH Oxidoreductases - biosynthesis, NADH, NADPH Oxidoreductases - genetics, Overexpression, Pyrococcus - enzymology, Pyrococcus - genetics, Pyrococcus furiosus, Recombinant Proteins - genetics, Recombinant Proteins - metabolism, TrpB gene, Tryptophan, Tryptophan - biosynthesis, Tryptophan synthase