title: Bacterial expression of two human aryl sulfotransferases.
abstract: The effect of replacing a single codon in the N-terminal of human aryl sulfotransferase (HAST) 1 and 3 with one that is more commonly found in E. coli genes was assessed. The pKK233-2 E. coli expression vector was employed and the polymerase chain reaction (PCR) was used to introduce the 5' nucleotide substitution, at the same time maintaining the fidelity of the amino acid sequence. The data indicates that this change had a minimal effect on protein production, subcellular localization or, in the case of HAST3, catalytic activity. In general, the pKK233-2 E. coli vector has been less than optimal for expressing human sulfotransferase cDNAs.
authors.affiliation: Department of Physiology and Pharmacology, The University of Queensland, Brisbane, Australia.
authors.forename: L M
authors.initials: LM
authors.lastname: Bidwell
authors.forename: E M
authors.initials: EM
authors.lastname: Gillam
authors.forename: A
authors.initials: A
authors.lastname: Gaedigk
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.forename: D
authors.initials: D
authors.lastname: Grant
authors.forename: M E
authors.initials: ME
authors.lastname: McManus
mesh_terms: D000818:Animals; D015239:Arylsulfotransferase / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D019556:COS Cells / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D011994:Recombinant Proteins; D015239:Arylsulfotransferase; D004298:Dopamine