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Assay of neomycin phosphotransferase activity in cell extracts

โœ Scribed by N. Ramesh; William R.A. Osborne

Book ID
102630516
Publisher
Elsevier Science
Year
1991
Tongue
English
Weight
293 KB
Volume
193
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

A simple method for the measurement of neomycin phosphotransferase (NPT) activity in crude extracts of eukaryotic cells is described. This method is based on the elimination of interfering phosphorylated proteins by using phenol-chloroform extraction. This solution phase assay allows the detection of greater than or equal to 0.01 ng of NPT in the crude cell extract. Rapid screening of a large number of cell cultures generated in gene-transfer experiments, using NPT as a selective marker, is made possible by this simple technique. Further, the promoter strength of vector constructs used in gene therapy may also be estimated by this procedure.

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โœ Steven G. Platt; Ning-Sun Yang ๐Ÿ“‚ Article ๐Ÿ“… 1987 ๐Ÿ› Elsevier Science ๐ŸŒ English โš– 874 KB

A dot assay for determining neomycin phosphotransferase (NPT II) activity in crude cell extracts has been developed. The assay provides for the rapid screening of large numbers of cell cultures generated in gene transformation experiments using NPT II as a dominant selectable marker. Currently, the