A structural view and understanding of NPF (nitrate transport) protein activity in maize

Summary

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.

Supervisor

Professor Brent Kaiser.

Research location

School of Life and Environmental Sciences

Program type

PHD

Synopsis

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 information

Additional supervisor for this project is Dr Chandrika Deshpande.

Expected Outcomes:
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:

• Confidential disclosure and registration of a disability that may hinder your performance in your degree;
• Confidential disclosure of a pre-existing or current medical condition that may hinder your performance in your degree (e.g. heart disease, pace-maker, significant immune suppression, diabetes, vertigo, etc.);
• Ability to perform independently and/or with minimal supervision;
• Ability to undertake certain physical tasks (e.g. heavy lifting);
• Ability to undertake observatory, sensory and communication tasks;
• Ability to spend time at remote sites (e.g. One Tree Island, Narrabri and Camden);
• Ability to work in confined spaces or at heights;
• Ability to operate heavy machinery (e.g. farming equipment);
• Hold or acquire an Australian driver’s licence; Hold a current scuba diving licence;
• Hold a current Working with Children Check;
• Meet initial and ongoing immunisation requirements (e.g. Q-Fever, Vaccinia virus, Hepatitis, etc.)

You must consult with your nominated supervisor regarding any identified inherent requirements before completing your application.

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Opportunity ID

The opportunity ID for this research opportunity is 2921

Other opportunities with Professor Brent Kaiser