LYMPHOCYTE PROTEINPAEDIA STAGE-2 - T-CELL POLYPEPTIDES FROM A PARTITIONED CDNA LIBRARY REVEALED BY THE DUAL DECAY METHOD

A complex population of gene products was analyzed by combining the gr eat resolving power of two-dimensional (2D) protein electrophoresis wi th a detailed dissection of individual protein species afforded by cDN A cloning. A cDNA library from BW5147 was partitioned by random sampli ng into sectors (of a complexity of 500 phase plaques/sector). From ea ch of the unique sets of cDNA clones present in the sector, 2D gels we re prepared. Patterns of spots were analyzed using the Kepler software system, and the compiled data both from the sectors and from the natu ral lymphocyte populations have been established to serve as a guide f or gene retrieval. Use of the dual decay method, which compares two ex posures of a 2D gel co-electrophoretic pattern, obtained from samples with (S-35)-polypeptides mixed with (H-3)-polypeptides, is described a nd scrutinized. It was determined that of 268 spots (three sectors com bined), 118 spots were shared with the natural lymphocyte population, the remaining 150 spots occur only in the sectors (and not detected in the cell population). All cDNA populations are available for retrieva l. Information obtained throughout the work was entered into the datab ase.