Cell-free translation of a chimeric eucaryotic-procaryotic message yields functional chloramphenicol acetyltransferase.
A vaccinia virus (VV) recombinant containing the DNA sequences encoding the bacterial chloramphenicol acetyltransferase (CAT) gene was constructed. The ability of the chimeric VV:CAT transcript to be translated in vitro into enzymatically active enzyme was assessed. Addition of mRNA isolated from the cytoplasm of VV:CAT infected cells to a mRNA-dependent reticulocyte lysate resulted in the synthesis of high levels of enzymatically active CAT. These results suggest that this assay may be used in concert with physical assays to study the expression and stability of chimeric transcripts in virus-infected cells.
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D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D002474:Cell-Free System; D015500:Chloramphenicol O-Acetyltransferase; D002855:Chromatography, Thin Layer / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005796:Genes; D005798:Genes, Bacterial; D005814:Genes, Viral; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D014616:Vaccinia virus / Q000235:genetics
D012333:RNA, Messenger; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase