Molecular cloning of cDNAs encoding two types of gonadotrophin α subunit from the masu salmon, Oncorhynchus masou: construction of specific oligonucleotides for the α1 and α2 subunits

Abstract

ABSTRACT Two types of cDNA (GTHα1 and -α2) encoding the α subunits of masu salmon (Oncorhynchus masou) gonadotrophin were cloned by the reverse transcription-polymerase chain reaction for pituitary mRNAs. The nucleotide sequences showed that the GTHα1 cDNA was 380 bp long, encoding 119 amino acids, and that GTHα2 cDNA was 365 bp long, encoding 114 amino acids. The masu salmon α subunit types had a few differences between the sequences, with homologies of 80% (nucleotide sequence) and 72% (amino acid sequence). The structural difference between the α1 and α2 subunits was predicted using hydropathic analysis. The evolutionary interval between masu and chum salmon was estimated to be 4·0 and 2·3 million years by comparing their GTHα1 and -α2 subunits respectively. These time values are roughly consistent with the evolutionary time interval (3·0 million years) estimated from fossil records and an isozyme study. Specific synthetic oligonucleotide probes were constructed and used for genomic Southern blot analyses. The restriction fragment sizes of the GTHα1 and -α2 genes were similar, and when their patterns were compared with those from four other teleosts, each species showed a different pattern from the others, but no difference between their respective α1 and α2 genes. Therefore, the structural features of the GTHα1 and -α2 genes may have diverged in a similar manner in these five teleosts.