Use of a cryptic splice donor site in the chloramphenicol acetyltransferase (CAT)-SV40 small-t antigen cassette generates alternative transcripts in transgenic rats.
The bacterial gene chloramphenicol acetyltransferase (CAT) is a widely used reporter in both in-vitro and in-vivo studies of genetic regulation. We have recently generated novel rat transgenic lines carrying an arylalkylamine N-acetyltransferase (AA-NAT) promoter-reporter construct in which CAT (with associated SV40 small-t antigen sequence) is the reporter. In addition to the predicted transgene transcript (1.9 kb), we identified an abundant 1.5 kb transcript which derives from an alternative splicing event that utilises a cryptic splice donor site located within the CAT gene. The native CAT open reading frame (ORF) is lost in the 1.5 kb transcript, and a western analysis has shown that protein deriving from an aberrant open reading frame is not expressed at detectable levels.
Burke
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School of Biosciences, Cardiff University, UK. smbdac@cardiff.ac.uk
Wells
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T
Carter
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Baler
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R
D017398:Alternative Splicing; D000818:Animals; D030801:Animals, Genetically Modified; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics* / Q000378:metabolism; D001191:Arylamine N-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics* / Q000378:metabolism; D005260:Female; D017930:Genes, Reporter; D008845:Microinjections; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes; D015053:Zygote
D000952:Antigens, Polyomavirus Transforming; D015500:Chloramphenicol O-Acetyltransferase; D001191:Arylamine N-Acetyltransferase