We propose to purchase a New Brunswick Scientific BIOFLO5000 60 liter working volume fermentor and a CEPA Z41 high capacity flow-through centrifuge to establish a Microbial Production Core Facility primarily for the growth of bacterial and yeast strains expressing recombinant proteins. The proposed core facility will serve the needs of 15 NIH-supported investigators, which are funded through 34 different NIH awards (22 R01, 1 R21, 1 R33, 2 R37 (MERIT), 2 P01, 1 P30, 1 P41, 1 S10, 1 T32, 1 U19, 1 U54). The core facility will overcome a significant bottleneck in protein purification for all laboratories involved. The investigators involved in this application work on 130 different proteins, often in their wild type and mutant forms, representing a total of over 200 protein preparations. The planned core facility is more efficient in growing large cultures than existing possibilities (10 liter fermentor, flasks), allows larger protein preparations, and will enhance the reproducibility of the starting cell material. Significant time savings for each participating research program will result. The leap in the amount of starting material will allow purifying preparative amounts of poorly soluble proteins at sufficient concentrations to finally allow their biochemical and biophysical analysis. Our choice of the New Brunswick Scientific BIOFLO5000 and the CEPA Z41 is based on previous experience with New Brunswick fermentors and CEPA products, the input from individuals running similar facilities, the pricing, and the specifications of the instruments. Relevance: The investigators of this submission engage in fundamental and applied biomedical research, using purified proteins to define mechanisms that are relevant to cancer and human health. The requested instrumentation will not only make their work significantly more efficient, but will also allow the analysis of proteins that previously could not be purified in sufficient quantities or concentrations. ? ? ?
| Liu, Jie; Doty, Tammy; Gibson, Bryan et al. (2010) Human BRCA2 protein promotes RAD51 filament formation on RPA-covered single-stranded DNA. Nat Struct Mol Biol 17:1260-2 |
