Expression of a human placental alkaline phosphatase gene in transfected cells: use as a reporter for studies of gene expression.
The human placental alkaline phosphatase gene has been cloned and reintroduced into mammalian cells. When a plasmid carrying the gene under control of the simian virus 40 early promoter (pSV2Apap) is transfected into a variety of different cell types, placental alkaline phosphatase activity can readily be detected by using whole cell suspensions or cell lysates. Alkaline phosphatase activity can also be visualized directly in individual transfected cells by histochemical staining. The gene is appropriate for use as a reporter in studies of gene regulation since its expression is dependent on the presence of exogenous transcription control elements. The overall assay to detect the expression of the gene is quantitative, very rapid, and inexpensive. Cotransfections of cells with pSV2Apap and a related plasmid carrying the bacterial chloramphenicol acetyltransferase gene (pSV2Acat) indicate that transcription of these two genes is detected with roughly the same sensitivity.
Henthorn
P
P
Department of Human Genetics, University of Pennsylvania School of Medicine, Philadelphia 19104-6072.
Zervos
P
P
Raducha
M
M
Harris
H
H
Kadesch
T
T
D000469:Alkaline Phosphatase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D005260:Female; D005796:Genes; D006801:Humans; D007700:Kinetics; D010920:Placenta / Q000201:enzymology*; D010957:Plasmids; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection
D000469:Alkaline Phosphatase