Relative promoter activity in human mammary epithelial cells assayed by transient expression.
Chimeric DNA expression vectors containing regulatory sequences proximal to the 5' end of coding sequences for mammalian genes provide valuable tools to study gene expression. Genes coding for easily measured products (reporter genes) can be used to study promoter strength and regulation of gene expression after transient expression of promoter-reporter constructs in mammalian cells. To determine the strength of a variety of mammalian and viral promoter-enhancer sequences in primary cultures of human mammary epithelial cells (HMEC), these sequences were fused to the bacterial chloramphenicol acetyltransferase (CAT) gene and transfected into HMEC using strontium phosphate. The long terminal repeat (LTR) of the endogenous murine leukemia virus AKR-623 was the most potent promoter of transient CAT expression in HMEC. A number of commonly available promoter sequences displayed a wide range of activities in these cells. The glucocorticoid responsive LTR promoter from the murine mammary tumor virus modulated expression of CAT and was sensitive to the concentration of dexamethasone in the growth media. In a similar fashion, the regulatory sequences from the murine metallothionein-1 gene retained responsiveness to zinc concentration in the growth media.
Huper
G
G
Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710.
Marks
J R
JR
Wiener
J R
JR
Iglehart
J D
JD
D000818:Animals; D001940:Breast / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology; D004247:DNA / Q000235:genetics; D003907:Dexamethasone / Q000032:analysis / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D015870:Gene Expression / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D015032:Zinc / Q000032:analysis / Q000494:pharmacology
D003470:Culture Media; D003907:Dexamethasone; D004247:DNA; D008668:Metallothionein; D015500:Chloramphenicol O-Acetyltransferase; D015032:Zinc