Application of the split-ubiquitin membrane yeast two-hybrid system to investigate protein interactions of small hydrophobic (SH) protein of human respiratory syncitial virus (HRSV).

• Main Author: Sailov, Talgat.
• Other Authors: Jaume Torres
• Format: Final Year Project
• Language: English
• Published: 2011
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/44754
• Institution: Nanyang Technological University

Human respiratory syncytial virus (HRSV) is a member of Paramyxoviridae family. It is a significant cause of lower respiratory tract infection in babies and children. There are no vaccines available for treatment of HRSV infection at the moment. Thus, the HRSV life cycle and pathogenesis is the subject of considerable research activity. Small hydrophobic protein (SH) is an integral membrane protein expressed in HRSV infected cells. Research shows that SH might play an important role in inhibiting TNF-alpha signaling. Thus, it is a potential drug target. The goal of the current project is to identify possible interaction partner of SH using yeast two-hybrid system. Because of hydrophobic nature of SH, its analysis is extremely difficult. Here, we used Membrane-based Yeast Two-Hybrid system to overcome this problem. Mainly, this assay utilizes separable domains of ubiquitin to study membrane protein interactions. In this project, we made a library screening of SH protein and tried to identify the proton conductance of SH by setting up the liposomal proton flux assay.