Tissue plasminogen activator (tPA) as a reporter gene in transient gene expression.
Using the gene coding for tissue plasminogen activator (tPA) as a reporter gene, a transient gene expression system has been established. Vectors containing the full-length cDNA of tPA with its signal sequences were introduced into mammalian recipient cells by a modified gene transfer procedure. Thirty hours after transfection, the secreted tPA was found in serum-free medium and measured by a fibrin-agarose plate assay (FAPA). In this assay, tPA converts plasminogen into plasmin which then degrades high-Mr fibrin to produce cleared zones. The sizes of these zones correspond to quantities of tPA. The combination of transient tPA expression system and the FAPA provides a quick, sensitive, quantitative and non-destructive method to examine the strength of eukaryotic regulatory elements in tissue-culture cells.
Cheng
S M
SM
Microbiology Division, Wyeth Laboratories Inc., Radnor, PA 19087.
Lee
S G
SG
Kalyan
N K
NK
McCloud
S
S
Levner
M
M
Hung
P P
PP
D000818:Animals; D002460:Cell Line; D004262:DNA Restriction Enzymes; D005786:Gene Expression Regulation; D005796:Genes; D005822:Genetic Vectors; D007739:L Cells / Q000378:metabolism; D051379:Mice; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D010959:Tissue Plasminogen Activator / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection
D004262:DNA Restriction Enzymes; D010959:Tissue Plasminogen Activator