Molecular cloning, nucleotide sequence and presence of multiple functional polyadenylation signals in the 3'-untranslated region of equine dopamine beta-hydroxylase cDNA.

Complementary DNA (cDNA) encoding equine dopamine beta-hydroxylase (DBH) was amplified with a combination of reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) method, and their nucleotide sequences (Accession No. AB029430: the DDBJ nucleotide sequence database) was determined. A total of 3842 bp cDNA sequence was consisted with 5 bp of 5' flanking untranslated sequence, 1833 bp of open reading frame encoding 610 amino acids, and 2004 bp of 3' flanking untranslated sequence. The deduced amino acid sequence of equine DBH was very similar to the known mammalian DBH sequences. The similarity between amino acid sequence of equine DBH to sequences of bovine, human, rat and mouse DBH were 86.3, 84.6, 82.2 and 81.2%, respectively. Northern blot analysis and in situ hybridization revealed three different sizes of mRNA expressions in equine adrenal medulla tissue. Then we found three putative polyadenylation signal sites in the 3' flanking untranslated sequence. These results indicate that alternative use of three polyadenylation sites generates the equine DBH mRNA that have different sizes of 3' flanking untranslated region. These results may provide further evidence for understanding DBH molecule and clues for the equine DBH gene analysis.

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