Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1996-12-06
1999-10-19
McKelvey, Terry
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 29, 435243, 435325, 435410, C12Q 168, C12Q 102, C12N 100, C12N 510
Patent
active
059687380
ABSTRACT:
Two spectrally distinguishable GFPs are used as reporters in mammalian cells to simultaneously and independently analyze the expressions of two transcriptional elements. The two GFPs, encoded by single stably integrated transcriptional elements, are readily and quantitatively detectable by FACS or flow cytometry. One of the GFP mutants (S202F, T203I, V163A) retains only the major excitation peak of wild-type GFP, while the other (S65T, V163A) retains only the minor excitation peak of wild-type GFP. Both variants have emission peaks overlapping that of wtGFP. The first mutant is excited at 406 nm using a Kr ion laser, while the second mutant is excited at 488 nm using an Ar ion laser. Emissions from both GFPs are measured at about 515 nm. The mutant excited at 406 nm can be used in conjuction with a fluorescein-based assay such as FACS-Gal. Applications include drug screening, measurements of temporal orders of gene expression, analysis of signal transduction pathways, and measurements of protein-protein interactions using two-hybrid systems.
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Anderson Michael T.
Herzenberg Leonard A.
McKelvey Terry
Sherwood Pamela
The Board of Trustees of the Leland Stanford Junior University
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