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University of Connecticut Health Center - Know Better Care The Gregory P. Mullen NMR Structural Biology Facility & Biophysical Core Facility

Protein Expression Service

CD

We can provide custom protein production of recombinant proteins suitable for biological assays and structural biology research. We can start with cloned proteins or we can assist in the cloning process to ensure that appropriate sequences, vectors and strains are being used.

As each system is different we start with a free consultation to develop an appropriate plan based on the researchers needs.

Bacterial Protein Expression

We can express proteins in bacteria in traditional flasks with shaking or with a New Brunswick BioFlo Fermenter capable of maintaining a controlled pH and with high levels of oxygenation. Expression can be performed on either rich media, for optimal expression, or minimal media for various labelling strategies needed in NMR and X-ray crystallography. Protein expression scales typically range from ug to mg quantities.

Protein Purification

We can provide cell pellets for the researcher to further purify or we can handle the protein purification. We can handle affinity purification, ion exchange chromatography, or size exclusion purification. We can also remove affinity tags if properly designed with a cleavage site.

Wheat Germ cell free expression

The facility has a Cell Free Sciences robot capable of performing Wheat Germ extract cell-free expression of affinity-purified protein in the 10 ug range. The robot is capable of performing 24 reactions in parallel, useful for exploring multiple constructs or increasing yield.

Pricing

Pricing for the protein expression and purification services is based on the number of hours of the technicians time plus any costs for supplies (e.g. media) and purchased services (e.g. DNA sequencing). As each system is different we do not provide a flat rate, but will provide a reasonably accurate quote during the free consultation.

Protein Characterization

In addition to protein expression and purification services we can provide characterization via CD to determine if the protein appears well folded and measures its thermal stability. Protein oligimerization states can be estimated from calibrated gel filtration chromatography or through analytical ultra-centrifugation performed at UConn Storrs at the National Analytical Ultracentrifugation Facility.