Postdoctoral Research Associate/Fellow
Click to apply Deadline: 10th January 2017
This Gordon and Betty Moore Foundation funded post is available from 1st February 2017 and based within Biosciences and the Living System Institute at the University of Exeter. This is part of a large project in collaboration with Prof. Thomas Richards at the University of Exeter and Dr. Alyson Santoro at the University of California, Santa Barbara. The successful applicant will investigate the function of membrane protein transporters from uncultured marine microbes by developing novel synthetic systems for studying substrate uptake kinetics in these transporters. The post will include working on model lipid bilayers (Cama et al. 2014, Lab Chip 14:2303 ; Cama et al. 2015, J Am Chem Soc 137:13836 ), transporter reconstitution, microfluidics (Pagliara 2014, Nat Mater 13:638) and quantitative image analysis. The successful applicant will be able to present information on research progress and outcomes, communicate complex information, orally, in writing and electronically and prepare proposals and applications to external bodies. Applicants will have a Ph.D. or equivalent in physics, chemistry, biology or biotechnology, demonstrate scientific excellence and have a strong publication record. Experience with model lipid bilayers, programming, image analysis and microfluidics is desirable.
EPSRC DTP PhD studentship: Developing novel optical and microfluidic tools to understand protein transporter function and kinetics
Click to apply Deadline: 11th January 2016
This project will develop a novel approach based on model lipid bilayers, microfluidics and microscopy to characterise transport across membrane protein pores and understand the physical mechanisms underlying substrate uptake in living systems. Dr Pagliara team recently used giant unilamellar vesicles (GUVs) and microfluidics to investigate molecular uptake through the outer membrane protein F, a transporter shown to play a role in bacterial antibiotic resistance (Cama et al. 2014, Lab Chip 14:2303 ; Cama et al. 2015, J Am Chem Soc 137:13836 ). Dr Petrov team have a strong track record in investigating elasticity of cell and model membranes (including GUVs) and the role of the membrane physical properties in biological function. Prof Richards team has optmised a protocol for purifying and tagging protein transporters to be reconstituted in GUVs. Building on this expertise, the student will obtain GUVs from lipids extracted from different microbial populations and reconstitute fluorescently labelled protein transporters in them. By engineering the geometry of bespoke microfluidic devices, the student will be able to trap GUVs embedded with transporters, deliver putative transported substrates and quantify substrate uptake and the number of reconstituted transporters via fluorescence microscopy. Quantitative image analysis and theoretical modeling (Pagliara et al. 2014, Phys Rev Lett, 113:048102; Locatelli et al. 2016, Phys Rev Lett, 117:038001) will allow determining substrate uptake kinetics, in terms of affinity, half-saturation constant and uptake rate, with single GUV resolution and comparing the different kinetics obtained for different combinations of protein transporters and GUVs. The effect of the reconstituted transporters on the mechanical properties of the membrane will be investigated using fluctuation spectroscopy and micropipette aspiration, techniques available in Petrov laboratory.
This project will help clarifying important biophysical questions related to the mechanisms of transmembrane transport in cells and could suggest novel approaches to designing antimicrobial drugs.
BBSRC SWBio DTP PhD studentship: Novel microfluidic technologies for single bacterium RNA-sequencing
Click to apply Deadline: 5th December 2016
Dr Pagliara laboratory has extensive experience in microfluidics (Pagliara 2014, Nat Mater 13:638) and is currently using the microfluidic mother machine technology
to image individual Escherichia coli bacteria and study the expression of selected genes of interest by using reporter-gene fusions. Prof John Love has extensive expertise in sorting single cells by using fluorescence activated cell sorting, Dr Thomas Connor is an expert in bioinformatics and bacterial genomics, Dr Konrad Paszkiewicz is an expert in RNA sequencing and transcriptome analysis.
You will build on this expertise to develop microfluidic devices capable of capturing single bacteria and prepare their lysate for off chip RNA-sequencing. This will allow us to obtain unprecedented understanding on the heterogeneity in gene expression in bacterial cells and how this links with cell-to-cell phenotypic differences in growth and resistance to stress, such as antibiotic exposure. You will receive a unique and comprehensive training including fabrication of devices with micrometer features, handling of microfluidics, fluorescence-activated cell sorting technologies, imaging via bright- field and fluorescence microscopy, programming image analysis software, handling microbial cultures, constructing reporter-gene fusions, extracting nucleic acids and preparing cDNA libraries, transcriptomic analysis and bioinformatics.