BACTERIAL EXPRESSION OF 2 HUMAN ARYL SULFOTRANSFERASES

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 intro duce the 5' nucleotide substitution, at the same time maintaining the fidelity of the amino acid sequence. The data indicates that this chan ge had a minimal effect on protein production, subcellular localizatio n or, in the case of HAST3, catalytic activity. In general, the pKK233 -2 E. coli vector has been less than optimal for expressing human sulf otransferase cDNAs. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.