This PhD project will develop and analyse high quality crystallised proteins of the nitrate/chloride transport proteins ZmNPF6;4, ZmNPF6;4 (Y370H) and ZmNPF6;6. The goal will be to identify the structural properties of these three membrane proteins to elucidate the location where either nitrate or chloride binds within their active pores and to resolve their functional activities in planta.
Full-length ZmNPF6;4 and ZmNPF6;6 cDNAs have been cloned and tagged with an in-frame C-terminal His (8) tag with a thrombin cleavage site and inserted into an expression vector pDEST8 (Thermo Scientific). Procedures to produce and purify ZmNPF6;6 and ZmNPF6;4 protein have previously been determined with pilot experiments at the University of Washington. Proteins will be overexpressed and purified using baculovar viruses that infect Sf9 insect cell lines within the Sydney Analytical – Protein Purification Facility. Concentrated ZmNPF6 proteins (10 mg/ml) will be crystalised using the hanging-drop-vapour diffusion method and analysed at the Australian Synchrotron facility at Monash University.
Additional supervisor for this project is Dr Chandrika Deshpande.
1) Development and analysis of crystal structures for the nitrate/chloride transport proteins ZmNPF6;4, ZmNPF6;4 (Y370H) and ZmNPF6;6.
2) Identification of nitrate and chloride binding sites within NPF proteins
HDR Inherent Requirements
In addition to the academic requirements set out in the Science Postgraduate Handbook, you may be required to satisfy a number of inherent requirements to complete this degree. Example of inherent requirement may include:
The opportunity ID for this research opportunity is 2921