Analysis of promoter and enhancer cell type specificities and the regulation of immunoglobulin gene expression.
We have analysed the properties of IgH promoter (VH) and enhancer (Ig) regions which were used to drive the expression of the chloramphenicol acetyl transferase (CAT) gene (cat) in recombinant plasmids. We observe little synergistic effect between the VH promoter and Ig enhancer on cat gene expression in our constructs. Replacing the VH promoter by the thymidine kinase (TK) promoter does not affect the enhancer-mediated B-cell-specific expression of the cat gene. However, replacement of the VH promoter by the mouse renin gene promoter, which is not normally expressed in B cells, completely abolishes cat gene expression in cells of this lineage. When the Ig enhancer is replaced by the SV40 enhancer (SV), CAT activity is restricted to B cells. The VH promoter is as efficient as the TK promoter in a preB cell line. Extending the size of the VH promoter fragment to include sequences between 126 to 639 bp upstream from the transcription start point results in an eight-fold decrease in CAT activity. In this situation, the tissue specificity of the promoter cat fusion is maintained. Among the various combinations tested here, the association of the TK promoter and the Ig enhancer expresses the cat gene most efficiently. The implications of these observations are discussed.
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology*; D015500:Chloramphenicol O-Acetyltransferase; D004742:Enhancer Elements, Genetic; D005347:Fibroblasts / Q000502:physiology; D005786:Gene Expression Regulation; D005809:Genes, Regulator; D007136:Immunoglobulins / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection
D007136:Immunoglobulins; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase