Project details – University of Copenhagen

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Understanding the transport mechanism of the NPF family

Main area:Plant biochemistry
Target group:Biology-Biotechnology, Natural Resources, Environmental Science
Educational level:Master, Bachelor, Project outside course scope
Project description:

Introduction

Transport engineering is an emerging research field in biotechnology with an immense potential within agro- and synthetic biology. To unleash its potential we must identify an arsenal of transport proteins with diverse substrate specificities and be able to engineer the properties of transport proteins. However, we lack basic understanding of the underlying transport mechanism of most transporters, which limits our ability to engineer fundamental properties such as substrate specificity, direction and speed of candidate transporters

Background

Our aim is to increase our understanding of the transport mechanism of one of the largest plant transport families, the nitrate and peptide transporter family (NPF). The NPF recently emerged as a family of transporters of phytohormones and specialized metabolites with high relevance for agro- and synthetic biotechnology. We have identified a powerful set of pairs of transporters similar in sequence but with distinct properties. Using these transporters we wish to elucidate the transport mechanism of the NPF family.

Perspectives

The immediate aim of the project is to increase our understanding of the transpot mechanism of the NPF. The long-term goal is to develop novel strategies for transport engineering within crop- and synthetic biology.

Expectations: We expect you to be enthusiastic, ambitious, hard working, dedicated and eager to learn and advance your skills in many disciplines within research and in oral and written presentations.

Who we are; Transport group of Hussam H. Nour-Eldin in the National Research Centre of Excellence, DynaMo (http://dynamo.ku.dk/). We provide a stimulating, international research environment where you have lots of opportunity to advance your skills. Our research lab has a strong international research profile with a recent record of high impact publications. We are currently focusing on elucidating the physiological role of phytohormone transporters, the transport mechanism of the NPF family and generating new transport engineering solutions for synthetic biology.

Methods used:The project will, depending on your interest, focus on identifying molecular determinants of one or two properties of the NPF family (e.g. substrate specificity, electrogenecity and/or transport rate). We have, so far, used Xenopus oocytes as expression system. However, we are hampered by low-throughput. In this project, you will setup human embryonic kidney cells and membrane potential sensitive dyes for high-throughput screening and generate and screen mutant libraries for altered transport properties. Homology modeling will put positive mutations into a structural context. In our work, we use molecular cloning, bioinformatics, heterologous expression in Xenopus oocytes, metabolite profiling, electrophysiology, homology modeling and molecular dynamics. The project provides a solid foundation for future work in both research labs and biotech industry, gives a solid foundation in basic skills used in molecular biology and an understanding of necessary bioinformatics tools.
Keywords:Plant biology-biotechnology, Transporter biology, NPF family, Phytohormone and specialized metabolite transport
Project home page: http://dynamo.ku.dk/
Supervisor(s): Hussam Hassan Nour-Eldin
Email:huha@plen.ku.dk