Application and limitations of differential hybridization in the isolation of T cell-specific cDNA clones.
We investigated the limitations and effectiveness of differential hybridization in the cloning of T cell-specific cDNA (complementary DNA) molecular clones. By using the technique with T cell and B cell cDNA probes, together with Northern blot analysis, we successfully isolated cDNA clones exclusively expressed in T cells from 1 X 10(4) plaque-forming units of a T cell hybridoma. These clones represent 0.068% of the mass of the cytoplasmic mRNA. Our result shows that differential hybridization is an effective procedure when used in combination with Northern blot analysis for screening of genes selectively expressed in T cells.
D000818:Animals; D002999:Clone Cells; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics*; D007425:Intracellular Membranes / Q000378:metabolism; D051379:Mice; D009693:Nucleic Acid Hybridization; D011132:Polyribosomes / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology*
D012333:RNA, Messenger; D004247:DNA