Konstruksi Mutan Protein Disulfida Isomerase pada Saccharomyces cerevisiae

Protein disulphide isomerase (PDI) is an enzyme that catalyses disulphide-bond formation in protein to maintain the native conformation, either through redox or isomerization reactions. Few PDI's have been isolated from various organism such as mammalian livers, plants, green algae and Saccharomyces cerevisiae. Disruption of the PDI in S. cerevisiae is haplo lethal indicating that the product of this gene is essential for viability. PDI consist of 1590 pb, but the essential domain on PDI gene have not been known complete. To study caracteristic PDI protein domains, in this research was undertaken construstion PDI mutant on S. cerevisiae. In vitro mutagenesis was carried out using hidroxilamin (HA) as mutagen. Recombinant plasmid were constructed by ligation of mutated DNA fragment (758 pb) into a vector (6300 pb) using T4 DNA ligase. The result showed some mutants were lethal, indicated that b and b' domains are essential for PDI on S. cerevisiae. Few mutants poorly growth, and the other showed well growth. Key words: construstion, protein disulfide isomerase, Saccharomyces cerevisiae