Detection of recombinant Pichia pastoris producing glargine

Abstract

Abstract The global prevalence of diabetes continues to rise. Diabetes management involves the administration of insulin to help regulate blood glucose levels. Currently, two main types of insulin are used for diabetes treatment: human insulin and insulin analogs. One such insulin analog is insulin glargine, categorized as a long-acting insulin. Four prominent organisms used for insulin production are Escherichia coli, Pichia pastoris, Saccharomyces cerevisiae, and Hansenula polymorpha. Pichia pastoris was used to produce insulin glargine in this study. The pPICZαA-G plasmid containing the synthetic glargine gene was inserted into Pichia pastoris. Recombinant Pichia pastoris containing the pPICZαA-G plasmid was detected using selection media with Zeocin, Polymerase Chain Reaction (PCR), and DNA sequencing. Through the implementation of selection media containing Zeocin, PCR, and DNA sequencing techniques, it was known that the recombinant Pichia pastoris contained the synthetic glargine gene. Further research will be carried out testing the expression of glargine in recombinant Pichia pastoris.