Expression of mouse::human immunoglobulin heavy-chain cDNA in lymphoid cells.
A chimeric mouse variable::human constant immunoglobulin heavy-chain gene was expressed in transfected mouse Sp2/0 cells. The chimeric immunoglobulin genes were integrated in tandem in the genome of stably transformed cells. These integrated gene copies were amplified by selection with a second drug marker. The gene amplification led to an increase in the expression of chimeric heavy-chain protein. The level of gene expression appears to be related to the site of integration; a few gene copies in one transfectant can yield as much heavy-chain protein as many copies in a second transfectant. In addition, we found that an adventitious oligo(C) sequence, introduced by our method of gene construction at a site located 8 nt residues downstream from a splice acceptor, can apparently direct splicing towards a cryptic splice acceptor downstream from the oligo(C).
D000818:Animals; D002460:Cell Line; D002678:Chimera; D004247:DNA / Q000235:genetics*; D005784:Gene Amplification; D005786:Gene Expression Regulation; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics*; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D010957:Plasmids; D014162:Transfection
D007143:Immunoglobulin Heavy Chains; D004247:DNA