Bovine Ornithine Decarboxylase Gene: Cloning, Structure and Polymorphisms

Abstract

Bovine ornithine decarboxylase (ODC) genomic clones were isolated from a bacteriophage lambda DASH genomic library. A total of 9452 bp sequence was determined which covers the entire sequence of the bovine ODC gene. Sequence analysis showed that the bovine ODC gene consisted of 12 exons which encode a protein identical to that inferred from a bovine ODC cDNA. Comparison of the structure and nucleotide sequence of the bovine, human and mouse ODC genes revealed that the gene was highly conserved. Primer extension analysis demonstrated that the transcription start point of bovine ODC mRNA was located 378 bp upstream from the A residue in the translation initiation codon. The 5'-untranslated region (UTR) of ODC mRNA was highly G + C rich, particularly in its 5'-most portion, and computer predictions suggested a very stable secondary structure for this region, with an overall free energy of formation of -134.4 kcal/mol. Conserved sequences and potential promoter elements including a TATA box, a possible CCAAT element, SP1 ranscription factor binding sites (GC boxes) and cAMP response elements (CRE) were identified in the 5'-flanking region of the gene. Two polymorphic restriction sites, a TaqI and a MspI, were mapped to the ODC gene and PCR-based methods for detection of the 2 polymorphisms were developed.