Open AccessOverexpression in Escherichia coli of the AT‐rich trpA and trpB genes from the hyperthermophilic archaeon Pyrococcus furiosus
Author(s) -
Ishida Masami,
Oshima Tairo,
Yutani Katsuhide
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb11433.x
Subject(s) - pyrococcus furiosus , chemistry , biology , microbiology and biotechnology , biochemistry , archaea , gene
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.