Recombinant Protein Expression and Molecular Cloning

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Each of the three laboratories involved in the Program Project will express distinct classes of proteins from serum proteins (ApoAI) to enzymes (LCAT, and MPO) and mutant forms of a subset of these in order to perform subsequent enzymatic assays as described in the Projects and analysis utilizing diverse and sophisticated instrumentation detailed in the Mass Spectrometry and Biophysics Core (Core B). The principal objective ofthe Recombinant Protein Expression and Molecular Cloning Core is to provide a centralized means for high volume protein expression thereby facilitating the relatively sophisticated and diverse biophysical and biological studies on the proteins of interest for each Project, in addition, the Core will also provide molecular cloning/ mutagenesis capabilities to all three Projects. The Core will also function as an educational facility for training members of the Program Project laboratories on ways to optimize protein expression using various strategies and also in the generation of mutants of specific alleles desired for protein expression or for targeting alleles for transgenic mouse production ( CC/CWRU Transgenic mouse Core is located at Case Western Reserve University). Training in the purification of proteins will also be provided by the Core. Centralizing these aspects of the work and training is expected to greatly enhance efficiency and productivity of the scientists participating in each project.