INTERAKSI IN VITRO ERF1 MUTAN Q359S TERHADAP ERF3 SACCHAROMYCES CEREVISIAE

• Main Author: Hermansyah
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/5259
• Institution: Institut Teknologi Bandung

<b>Abstrct:<p align="justify"> <br /> <br /> <br /> eRF1 and eRF3 proteins were believed to be involve in translation termination process. These proteins could interact each other to form functional complex of release factors, however detail interactions <br /> a of both proteins are needed to be elucidated. The C-terminal region of eRF1 was suggested as an essential domain for its interaction with eRF3. In this research in vitro interaction between Q359S eRF1 with eRF3 was carried out. pITB53 plasmid which carrying sup45-11s gene was used to express Q359S eRF1 mutant protein in Escherichia coll. SDS-PAGE analysis of E.coli cell free extract which harbouring pITB53, showed an overexpression pattern (44,95%) of mutant protein . Furthermore, this protein has been purified by using affinity chromatography IMAC system with Talon resin. Crude extract of eRF3 protein was prepared from S.cerevisiae , Al F2(803) which carrying pUKC606. The purified eRF1 and crude extract of eRF3 was used to in vitro interaction assay by using IMAC system. The result showed that Q359S eRF1 has higher binding affinity than that eRF1 wild type.