The molecular biology and protein expression core is organized on the basis of the pooling of the resources and expertise of all four project leaders in the program. This centraliz&l resource provides service and assistance in molecular biology, particularly the ability to rapidly subclone the gene of interest or a mutagenized derivative into one or more expression vectors without the need for restriction digestion. The core also provides the facilities and hardware for mammalian expression, selection of stable cell lines and large scale cell growth for collection of serum-free conditioned media. The core will provide facilities for large scale isolation of recombinant proteins, the appropriate chromatography strategies and will maintain the affinity columns necessary for this activity. Assistance and facilities will also be provided for the large-scale isolation of coagulation proteins from plasma. Finally, the core will provide some quality control services particularly the quantitation of 7 carboxy-glutamic acid content to verify the integrity of expressed vitamin K- dependent proteins. The Co-Directors will be responsible for the oversight and efficient conduct of these activities and for ensuring equitable service to the projects.
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