Kasha Laboratory | Institute of Molecular Biophysics
Protein Expression Facility

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Protein Expression Facility (PEF)

The PEF is a multi-user facility which serves primarily faculty and students from the laboratories of Structural Biology in the Kasha Laboratory Building. In addition laboratories in Biology, Biochemistry, Chemistry, Nutrition perform projects in the Facility.

Types of projects

The primary purpose of the PEF is to facilitate the large scale expression of recombinant proteins in eucaryotic cells. We have recently acquired new methods and equipment for that purpose, including the WAVE bioreactor and the BelloCell bioreator. Other projects include expression of native proteins in response to chemical or biochemical inducers or metabolic pressures, uptake of nanoparticles and dyes, production of siRNA knockout gene clones and analysis of effects, and cytological studies of the cellular location of novel proteins.

Special techniques and capabilities

Expertise includes growth of virus with host cells, both insect (Baculovirus) and mammalian (SV40, adenovirus, AAV), isolation and purification of the viruses, immunoassay development, transient transfection of mammalian cells, and selection and isolation of stable cell clones; growth assays for measurement of stimulatory effects of factors; cell analysis (viable cell density, cell size, cell shape) using a CedexHiRes. Most techniques involving cultured eucaryotic cells can be handled in the facility.

Protein Expression Facility Fee Structure

In November 2009 all of the IMB/KLB Core Facilities will begin assessing fees to partially defray the cost of the Facilities. The fees for PEF represent only the actual cost of materials. The Actual PEF User Fees can be consulted for various types of work to be performed at the Facility.

Initiating a project in the PEF

The process begins with an initial meeting with PEF director involving the principal investigator and the individual who will perform the work in the facility. A plan will be laid out for the scope and extent of the proposed work and an order form for the work will be filed. This will allow the facility to have adequate materials on hand for the project and provide documentation for the end of the month billing. A training schedule for the worker will be negotiated at the same time.

Working in the Facility

The lab is a biosafety level 2 (BSL2) laboratory designated by FSU Environmental Health and Safety, according to the CDC/NIH Joint Guidelines on Biosafety in the Biomedical Laboratory. The suite has been designed specifically to facilitate cell culture projects by multiple users- in both culture rooms which are isolated from all others and in common areas, such as the microscopy area and analytical area where users can gather to compare and discuss results. In addition to the entire suite being isolated, each culture room is also isolated containing all necessary equipment and supplies for culture within the room. Four biosafety cabinets continuously filter the air; six incubators allow isolation of cultures by bio-safety level. A manual of biosafety rules and regulations guides behavior and introduces users to biosafety equipment and design.