A method to detect transfected chloramphenicol acetyltransferase gene expression in intact animals
- Memorial Sloan-Kettering Cancer Center, New York, NY (USA)
A rapid procedure is described for assaying chloramphenicol acetyltransferase enzyme activity in intact animals following transfection of the RSV CAT plasmid into mouse bone marrow cells by electroporation. The reconstituted mice were injected with ({sup 14}C)chloramphenicol and ethyl acetate extracts of 24-h urine samples were analyzed by TLC autoradiography for the excretion of {sup 14}C-labeled metabolites. CAT expression in vivo can be detected by the presence of acetylated {sup 14}C-labeled metabolites in the urine within 1 week after bone marrow transplantation and, under the conditions described, these metabolites can be detected for at least 3 months. CAT expression in intact mice as monitored by the urine assay correlates with the CAT expression in the hematopoietic tissues assayed in vitro. This method offers a quick mode of screening for introduced CAT gene expression in vivo without sacrificing the mice.
- OSTI ID:
- 5451423
- Journal Information:
- Experimental Cell Research; (United States), Vol. 174:1; ISSN 0014-4827
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CARBON 14 COMPOUNDS
AUTORADIOGRAPHY
TRANSFERASES
ENZYME ACTIVITY
BONE MARROW CELLS
CHLORAMPHENICOL
GENE REGULATION
GENES
MICE
PLASMIDS
THIN-LAYER CHROMATOGRAPHY
URINE
ANIMAL CELLS
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BIOLOGICAL MATERIALS
BIOLOGICAL WASTES
BODY FLUIDS
CARBON COMPOUNDS
CELL CONSTITUENTS
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
DRUGS
ENZYMES
LABELLED COMPOUNDS
MAMMALS
MATERIALS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
VERTEBRATES
WASTES
550201* - Biochemistry- Tracer Techniques