We propose to set up a Molecular Resources Core Laboratory (MRCL) to fully support the methodologic requirements of the scleroderma SCOR. Thus, we envision MRCL as an efficient and economical facility equipped with specialized instruments and expertise to generate key reagents, and to continue to engage in developing state of the art techniques to facilitate the specific aims of the various sub-components of the scleroderma SCOR. The MRCL will be responsible for the purification, characterization and quality control of types I and III collagen and production and purification of cyanogen bromide sub-fragments of type I and III collagen. The Core will help design recombinant DNA vectors that are optimally suited to produce large amounts of wild type and site-specifically mutated 90-kDa receptor used for types I and III collagen binding on platelets. Furthermore, the MRCL personnel will collaborate with the leaders of the individual sub-projects of scleroderma SCOR to carry out the nucleic acid manipulations needed to quantify expression of multiple gene families. Finally, the MRCL will develop expression vectors that may be employed to produce dominant negative proteins (e.g., Smads) or antisense oligoribonucleotides in various types of mesenchymal cells. Thus, MRCL is proposed not only as a centralized resource for specialized biological reagents and equipment, but housed with a staff who will keep abreast of the newer experimental approaches of cell and molecular biology, deemed to be instrumental in successfully accomplishing the mission of the scleroderma SCOR.
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