Figure 1
Cloning of the two mouse peroxisomal thiolase genes and determination of the transcription initiation sites of each gene. a) Southern blot of a 129SV mouse genomic DNA digested with different restriction enzymes. Membrane was hybridized with an exon 9 rat thiolase 32P-labelled cDNA probe. A molecular weight marker DNA is indicated on the left in kb. b) Structural organizations of the mouse thiolase A and B genes: exon-intron distribution. Exons are numbered from I to XII. Translated sequences are shown as black boxes and untranslated sequences are open boxes. c) Identification of the transcription initiation sites of mouse thiolase A and B genes by ribonuclease protection assay. RNA probes corresponding to the genomic sequence extending upstream from the exon 1-intron 1 border (probes size: 678 nt and 521 nt for mThA and mThB, respectively) were hybridized to mouse liver RNA (lanes 1 and 6) and digested with RNAses. The protected fragments were analysed on a denaturing polyacrylamide gel (for details, see Methods). Lanes 2–5 and 7–10, DNA sequencing ladder from plasmids containing +1 mThA and +1 mThB probes, respectively. Arrows and dots indicate the different bands corresponding to multiple transcription initiation sites.