KARAKTERISASI KANDIDAT MUTAN HASIL MUTASI ACAK PADA DAERAH HPAL-BGILL GEN PDII SACCHAROMYCES CEREVISIAE

• Main Author: Aznury (NIM : 20596507), Martha
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/4808
• Institution: Institut Teknologi Bandung

<b>Abstract :</b><p align=\"justify\">Protein disulfide isomerase (PDI) is an enzyme involved in the native disulphide bond formation of secretory proteins. PDI had been isolated and characterized from prokaryote and eukaryote. In yeast Saccharomyces cerevisiae, PDI is encoded by PDII gene. Yeast PDII have two domains containing active sites (domain a and a\') and two putative binding sites domains (domain b and b\'). This research was conducted to analyze phenotype and genotype of four candidate mutants (yITB2, yITB4, yITB6 and yITB7) previously generated from random in vitro mutagenesis of the Hpal-BgIlI region of PDII. The yITB2 and yITB4 candidate mutants showed lethal phenotype while the yITB6 and yITB7 candidate mutants appeared to be more sensitive to DTT compared to that of the wild type yeast. However there were no mutations found in the Hpal-BglII regions of those four candidate mutants. The lethal phenotype shown by yITB2 and yITB4 was resulted from high stability of pCT37 carried PDII wild type and URA3 genes which could not be removed by growing on the non-selective medium for 50 generations. Hence, the lethal phenotype was due to the presence of URA3 gene product which converted 5\'-fluoroorotic acid (FOA) into toxic intermediate. The DTT sensitivity shown by yITB6 and y!TB7 might be caused by different physiology possed by individual yeast colonies.