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Heterologous production of hPTH in fission yeast Schizosaccharomyces pombe

Nipunjot K. Soni and Praveen P. Balgir

Human parathyroid hormone (hPTH) is a peptide hormone consisting of 84 amino acids (hPTH(l-84)). A gene coding for human parathyroid hormone (hPTH) was synthesized and cloned into REP1 expression vector. REP1 is an expression vector which has nmt1 strong promoter which is thiamine repressible. The DNA insert was designed for secretory production of Human Parathyroid hormone by linking cpy1 secretory signal sequence at 5’ end of cDNA sequence of Human Parathyroid hormone so that the expressed protein is excreted to the growth medium, allowing for rapid and easy purification of the desired products. The REP1 vector containing the designed DNA insert was transformed into Schizosaccharomyces pombe FY12854 strain. The transformed host was grown in Essential Minimal Media and extracellular media was checked for production of hPTH. Secreted protein was concentrated and purified by passing through S Sepharose column and further purified by two phase HPTLC.