Metallothionein-I promoter-directed expression of foreign proteins in a mouse pituitary corticotrope tumor cell line.
Expressing foreign proteins in heterologous eukaryotic cells has been a powerful tool for analyzing protein structure and function. The inducible mouse metallothionein-I promoter has been particularly useful for expression studies. However, the levels of expression achieved with this promoter in heterologous eukaryotic expression systems have not equaled those observed in vivo for the metallothionein-I gene. We have constructed expression plasmids placing either the gene for chloramphenicol acetyltransferase (CAT) or the cDNA for human neuropeptide Y (NPY) under control of the mouse metallothionein-I promoter. These two expression vectors were used to transfect mouse anterior pituitary tumor cells, from which stable transformants were isolated. The resulting cell lines, Mt.NPY1a and Mt.CAT, were used to maximize functional product expression from the metallothionein-I promoter. In both cell lines, a 35-fold induction of mRNA accumulation, peptide synthesis, or CAT activity was observed.
Dickerson
I M
IM
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Peden
K W
KW
Mains
R E
RE
D000818:Animals; D002104:Cadmium / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation; D008668:Metallothionein / Q000235:genetics*; D051379:Mice; D009478:Neuropeptide Y / Q000096:biosynthesis / Q000235:genetics; D010903:Pituitary Gland, Anterior / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured; D015032:Zinc / Q000494:pharmacology
D009478:Neuropeptide Y; D011993:Recombinant Fusion Proteins; D002104:Cadmium; D003907:Dexamethasone; D008668:Metallothionein; D015500:Chloramphenicol O-Acetyltransferase; D015032:Zinc