Structure of the channel catfish (Ictalurus punctatus) growth hormone gene and its evolutionary implications

A DNA fragment of 1.6 kilo base pairs (kb), encoding part of the channel catfish (Ictalurus punctatus) growth hormone (GH) gene, was generated by the polymerase chain reaction (PCR) using 2 degenerate synthetic oligonucleotides (30 and 33 mer) derived from the N- and C-terminal amino acid sequences of the catfish GH polypeptide as amplification primers and with catfish genomic DNA as a template. This DNA fragment was used as a probe for the isolation of a catfish GH gene from a genomic library constructed in a lambda phage cloning vector, lambda Dash II. Three positive clones were isolated, and their complete nucleotide sequences were determined. Nucleotide sequences from clones 1 and 3 were identical, whereas clone 2 had 2 base substitutions. The gene spans approximately 3 kb and is comprised of 5 exons and 4 introns. The initiation codon, the termination codon, and the canonical polyadenylation sequence were identified. The amino acid sequence deduced from the predicted coding region of the gene is in agreement with that of the native GH polypeptide sequence. A sequence (TATAAAA) matching the TATA box consensus sequence was located at nucleotide positions -30 to -23. Furthermore, 2 sequences corresponding to the mammalian Pit-1/GHF-1 binding sites (consensus sequence TT[AA]TATNCAT) were identified in the 5' flanking region starting at positions -113 and -134. Another sequence (GTACCAGTGA) conserved among the GH genes of the channel catfish and other known animal species was also identified at position -220. The biological functions of this sequence remain to be determined.(ABSTRACT TRUNCATED AT 250 WORDS)