A method to detect transfected chloramphenicol acetyltransferase gene expression in intact animals.
A rapid procedure is described for assaying chloramphenicol acetyltransferase (CAT, EC 2.3.1.28) 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 [14C]chloramphenicol and ethyl acetate extracts of 24-h urine samples were analyzed by TLC autoradiography for the excretion of 14C-labeled metabolites. CAT expression in vivo can be detected by the presence of acetylated 14C-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.
Narayanan
R
R
Memorial Sloan-Kettering Cancer Center, New York, New York 10021.
Jastreboff
M M
MM
Chiu
C F
CF
Ito
E
E
Bertino
J R
JR
D000123:Acetyltransferases / Q000032:analysis / Q000235:genetics*; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001853:Bone Marrow / Q000201:enzymology; D002701:Chloramphenicol / Q000652:urine; D015500:Chloramphenicol O-Acetyltransferase; D005796:Genes; D051379:Mice; D008822:Mice, Transgenic; D014158:Transcription, Genetic; D014162:Transfection
D002701:Chloramphenicol; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase