Life Sciences & Biotechnology
Title : | An integrated structural biology approach to unravel a unique fatty acid activation mechanism in eukaryotes |
Area of research : | Life Sciences & Biotechnology |
Focus area : | Fatty acid activation mechanism in eukaryotes |
Principal Investigator : | Dr Rajan Sankarnarayanan, Scientist, CSIR-Centre for Cellular and Molecular Biology (CSIR-CCMB), Hyderabad |
Timeline Start Year : | 2020 |
Timeline End Year : | 2025 |
Contact info : | director@ccmb.res.in |
Details
Executive Summary : | Objective: Generating loss of function mutation in Yeast to study the physiological role of Dip2 .Generating loss of function mutation and overexpression lines in Drosophila .Generating loss of function mutation in embryonic stem cells .Expression profiling to identify the stages for enhanced expression to obtain quality cDNA . Expression and purification optimization in BL21(DE3), Pichia and Sf9 expression systems . Biochemical characterization of various proteins . Identification of various phenotypes in loss of function mutants along with effects of different stresses . Complementation and Localization studies . Screening for Crystallization conditions for various constructs and complexes . Complementary structural characterization using SAXS, EM . Obtain quality preparative samples for performing proteomic studies after proximity labelling Obtain quality preparative samples for performing lipidomic studies and preliminary characterization of lipid preparations using thin layer chromatography . Separation of soluble and membrane protein extracts on gel electrophoresis Performing MS and MS/MS analysis to identify interacting proteins. Performing lipidomics in Multiple reaction monitoring (MRM)-mode and discovery- mode to identify the range of lipids. |
Organizations involved