Project details – University of Copenhagen

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Single-cell proteomics approach to identify plant “gatekeeper”

Main area:Molecular Biology
Target group:Biology-Biotechnology, Agriculture, Other Education
Educational level:Bachelor, Master, Project outside course scope
Project description:

The goal of this project is to identify and characterize the ‘gatekeeper’ that is key for driving the glucosinolate defense compounds into their storage site, the S-cells. Understanding this process will advance basic knowledge on how defense compounds move to sink tissue for optimal plant defense.  

Background: In the model plant Arabidopsis, glucosinolates are stored to high concentrations (>130 mM) in the vacuole of the 1 mm long laticifer-like sulfur-rich cells called the S-cells. An energy-requiring importer is responsible loading the glucosinolates into the vacuole. Single-cell sampling of the S-cells is feasible, which enables single-cell proteomics on the S-cells. This will provide a representation of the proteins expressed in the S-cells, including candidate vacuolar importers.

Activities:

a)     Single-cell proteomics on S-cells to identify candidate transporter

b)    Clone candidate transporter into yeast and plant expression vector

c)     Biochemical characterization of transporter in yeast (e.g. substrate specificity, Km value) and in planta (localization, expression level)

d)    In planta validation with mutants, metabolite profiling and bioassay

Work environments: We expect you to be enthusiastic, dedicated and eager to learn and advance your skills in many disciplines (e.g. research, oral and written presentations). In return, we provide a stimulating, international research environment, where you get lots of opportunity to advance your skills within basic plant and yeast methodologies.  

Methods used:molecular biology, heterologous expression in yeast and plant hosts, biochemical characterization of transporters, LC-MS analytical chemistry, transport engineering, plant genotyping, bioinformatics on candidate gene family.
Keywords:Proteomics, Transport biology, Yeast, Plant, Metabolite profiling
Project home page: http://dynamo.ku.dk/
Supervisor(s): Main supervisor: Barbara Halkier, Co-supervisor: Deyang Xu, dyxu@plen.ku.dk
Email:bah@plen.ku.dk