CHARACTERIZATION OF UPSTREAM REGION (5’UTR) OF THE GPDH3 GENE AS A PROMOTER IN CHLAMYDOMONAS REINHARDTII MICROALGAE

• Main Author: Pratiwi, Delfianti
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/54990
• Institution: Institut Teknologi Bandung

Chlamydomonas reinhardtii microalgae is one of promising models of organisms for industrial worlds to produce various chemical compounds and proteins, due to its rapid growth time, simple cultivation process, and established genetic modification. C. reinhardtii can produce proteins in an inducible and constitutive ways. Expression systems with inducible promoters have advantages over constitutive promoters, since the expression of recombinant proteins can be controlled at any time using certain inducers. C. reinhardtii has a glycerol phosphate dehydrogenase (CrGPDH3) gene where the upstream part acts as a promoter induced by NaCl. If this promoter can be isolated and used for production of recombinant proteins in microalgae, then the production cost of such proteins will be relatively cheap. The purpose of this study was to characterize the upstream area of the 5'UTR CrGPDH3 gene as a potential promoter in the microalgae C. reinhardtii. To achieve this goal, nucleotide sequences homology of CrGPDH3 and GPDH (Glycerol-3-phosphate dehydrogenase) genes derived from several organisms were analyzed using ClustalW and MEGA X software. In this study, characterizations of the upstream section of CrGPDH3 (PromA) were also conducted with the PlantCare, PlantPAN, PLACE, and Softberry database. Homological analysis showed that CrGPDH3 shared the highest homology of GPDH genes from Volvox carteri with a percentage of identity of 61.45%. Characterization of PromA of CrGPDH3 indicates the presence of the promoter main elements on this nucleotide sequence, including TATA box, CAT box, CAAT box, and CCAAT box. In addition, five elements of cis regulators were found, namely ABRE, DRE, MYB, MYC, and TGACG motif (MeJA motif) that allegedly regulate expression systems in the PromA promoter of C. reinhardtii under high salinity conditions. Based on homology analysis that has been done, several candidates for primer pairs have been successfully designed for future use in isolating potential promoters from C. reinhardtii. Therefore, PromA is likely to be a potential promoter to express genes encoding certain proteins in the C. reinhardtii microalgae system.