Expression of human thyroid peroxidase in the yeasts Saccharomyces cerevisiae and Hansenula polymorpha.
Saccharomyces cerevisiae and the methylotrophic yeast Hansenula polymorpha have been used to express both full-length and a large hydrophilic domain of human thyroid peroxidase (TPO). Expression of TPO in S. cerevisiae, using the natural signal sequence or the yeast alpha-mating factor (MF alpha) signal sequence, resulted in undetectable or very low levels of recombinant TPO production. However, TPO was expressed when the natural TPO leader sequence was replaced by the yeast STE2 signal sequence. This recombinant TPO reacted with both rabbit anti-human TPO polyclonal and mouse anti-human TPO monoclonal antibodies on Western blots. In the case of H. polymorpha, TPO expression was achieved when the natural TPO leader sequence was replaced by the MF alpha leader and the construct placed under the control of the methanol-regulated promoter from the methanol oxidase gene. The recombinant TPO produced in H. polymorpha reacted with both TPO polyclonal and TPO monoclonal antibodies. No TPO was produced when the signal sequence of SUC2 (invertase) or the TPO natural signal sequence was used to direct expression.
Department of Medicine, University of Wales College of Medicine, Heath Park, Cardiff, UK.
N
N
Wedlock
J
J
Furmaniak
S
S
Fowler
Y
Y
Kiso
J
J
Bednarek
A
A
Baumann-Antczak
C
C
Morteo
P
P
Sudbery
A
A
Hinchcliff
B
B
Rees Smith
D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D004247:DNA; D007453:Iodide Peroxidase