Rapid and Accurate Phenotype Screening at the Molecular Level will Play a Crucial Role in Mammalian Functional Genomic Research (February 2000)

Using the mouse as a model, we are currently developing a procedure that combines the phenotype- and gene-driven approach to mutation analysis. The first series of mutation studies involves mouse hemoglobin protein variant identification. Our preliminary studies which employed a novel rapid mass spectrometric separation and analysis scheme (on purified human hemoglobin), resulted in a recent accelerated publication in Analytical Chemistry (2000, 72, 899-907) entitled: "Ion Trap Collisional Activation of the (M+2H)2+ - (M+17H)17+ Ions of Human Hemoglobin b-Chain". This methodology allowed for separation times on the millisecond time scale, while minimizing the sample preparation steps prior to analysis. Recently, we have begun analyzing hemoglobin variants from the reference CH3 mouse strain and the C57BL/6 black mouse strain. Preliminary results suggest that differentiation of minor expression products from heterozygous mice are possible using newly developed mass spectrometric techniques described in the aforementioned paper (see figure below). Work in the next several months will focus on the bioinformatics associated with these types of measurements. . (Funding Source: DOE-OBER and LDRD; Contacts: Jim Stephenson, CASD, 574-2848 or stephensonjl@ornl.gov ; Michelle Buchanan, LSD, 574-4521 or buchananmv@ornl.gov; Eugene Rinchik, LSD, 241-2158, or rinchikem@ornl.gov)