The overall goal of the Transgenic Animal/Recombinant Protein Core Laboratory is to undertake and coordinate the breeding/cross-breeding of transgenic mice, and to prepare large amounts or recombinant proteins (sRAGE, sVCAM,-1, and RAGE-poliovirus receptor chimeras) for use in the proposed studies. Mouse breeding takes place in the Institute of Comparative Medicine of Columbia University, on the eighteenth floor of the P&S buildings (one floor above the laboratories where all three projects will be performed). Preparation of recombinant proteins is undertaken in laboratories of the investigators on the PPG) where there are tissue culture facilities for growing Sf9 cells and for performing infections with baculovirus constructs, as well as equipment for protein purification on an analytic and preparative scale (HPLC, FPLC, Isoelectric focussing, Smart and conventional chromatographic systems). Drs. Schmidt and Costantini, with the assistance of Dr. Yemul, will be responsible for breeding of transgenic mice. Dr. Schmidt, who discovered RAGE, is an expert in protein chemistry and will also supervise Dr. S. Yemul in the expression and purification of sRAGE, sVCAM-1 and related molecules. Each will be produced using the baculovirus system, and purified by methods which Dr. Schmidt has already worked out. In addition, Drs. Schmidt and Yemul will prepare and characterize AGE-modified proteins for studies proposed in the projects. This Core Laboratory will be utilized by each of the three components of the PPG.
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