Nonchromatographic assay for expression of the chloramphenicol acetyltransferase gene in eukaryotic cells

Journal Article · Wed Jan 01 00:00:00 EST 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90180-6· OSTI ID:6151582

Sleigh, M J

A rapid procedure is described for assaying chloramphenicol acetyltranserase (CAT) enzyme activity following transfection of the CAT gene into eukaryotic cells. CAT enzyme activity in cell extracts catalyzes the transfer of (/sup 14/C)acetyl groups from labeled acetyl coenzyme A to unlabeled chloramphenicol. Labeled reaction product is quantitated by liquid scintillation counting after extraction into ethyl acetate. The method is valid for use with transfected cell extracts only if the extracts are first heated to 65/sup 0/C to remove a factor which degrades acetyl coenzyme A. The revised procedure offers considerable advantages in speed and ease of performance over the chromatographic assay in current use.

• Research Organization: CSIRO, New South Wales (Australia)
• OSTI ID: 6151582
• Journal Information: Anal. Biochem.; (United States), Vol. 156
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
TRANSFERASES
BIOCHEMICAL REACTION KINETICS
GENES
ANIMAL CELLS
CARBON 14
CHLORAMPHENICOL
COENZYMES
LABELLED COMPOUNDS
TRACER TECHNIQUES
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON ISOTOPES
DRUGS
ENZYMES
EVEN-EVEN NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
NUCLEI
RADIOISOTOPES
REACTION KINETICS
YEARS LIVING RADIOISOTOPES
550401* - Genetics- Tracer Techniques