Expression of the Thy-1 glycoprotein gene by DNA-mediated gene transfer.
We isolated a gene encoding the Thy-1.2 glycoprotein from a recombinant library constructed from BALB/c mouse DNA. To evaluate the expression of this cloned gene in different genomic environments, we introduced it into cell lines derived from fibroblast, lymphoid, and neuronal tissues by DNA-mediated gene transfer. When integrated into the genome of mouse L cells, cell-surface Thy-1 can be detected with anti-Thy-1 monoclonal antibodies. These L-cell lines contain between two and four copies of the cloned Thy-1 gene stably integrated in the host genome. After subcloning into a plasmid vector containing the bacterial Eco-gpt gene as a selectable marker, the Thy-1 gene was introduced into the Thy-1-deficient mouse lymphoma AKR1 (Thy-1-d), and the rat neuronal cell line, B50. The resulting transformants also contain two to four copies of the cloned Thy-1 gene but express up to 50-fold more cell-surface Thy-1.2 than the L cell transformants. The expression of vastly differing amounts of cell-surface Thy-1 from similar numbers of genes suggests that the gene encoding this differentiation antigen is under tissue-specific regulation.
D000818:Animals; D000954:Antigens, Surface / Q000235:genetics*; D001483:Base Sequence; D001921:Brain; D002460:Cell Line; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism*; D005796:Genes; D007739:L Cells / Q000276:immunology; D007942:Leukemia, Experimental / Q000276:immunology; D051379:Mice; D010954:Plasmacytoma / Q000276:immunology; D010957:Plasmids; D018800:Thy-1 Antigens; D014162:Transfection
D000954:Antigens, Surface; D004274:DNA, Recombinant; D018800:Thy-1 Antigens; D004262:DNA Restriction Enzymes