Detection of the neor gene expression in animal cells after genetic transformation.
A reliable assay is reported for the detection of the marker gene aminoglycoside phosphotransferase activity in cells that express this enzyme transiently or as a result of stable genetic transformation. This method combines the simplicity of the dot assays with the reliability of the more elaborate and time consuming electrophoretic or chromatographic methods. Inhibition of phosphatases and protein kinases during the reaction reduces labeled ATP consumption by these enzymes. As a result, this assay allows the detection of approx 10 times lower levels of the enzyme than currently used methods. To detect the expression of reporter genes in transformed cells aminoglycoside phosphotransferase can be used as well as the widely used chloramphenicol acetyltransferase enzyme.
Szelei
J
J
Institute of Biochemistry, MTA Biological Center, Szeged, Hungary.
Szlanka
T
T
Duda
E
E
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D005189:False Positive Reactions; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D019868:Kanamycin Kinase; D010770:Phosphotransferases / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D015500:Chloramphenicol O-Acetyltransferase; D010770:Phosphotransferases; D019868:Kanamycin Kinase