Production of a recombinantly derived growth hormone antibody and the characterization of growth hormone levels in yellow perch
2004
Roberts, S. | Barry, T. | Malison, J. | Goetz, F.
Because of restrictions on commercial fishing and declining wild populations, there has been increased interest in the aquaculture of yellow perch. However, the slow growth of yellow perch has been an impediment to the development of this species for aquaculture. Using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), a growth hormone (GH) cDNA was isolated from yellow perch pituitaries that was 935 bp, coding for a protein of 204 amino acids. Yellow perch GH is most similar to sea bream GH at the amino acid level (88% identical). The cDNA was used to produce a recombinant protein in the pBAD TOPO vector, and the protein was used as an antigen to generate a polyclonal antibody to yellow perch GH. The antibody recognizes a 21-kDa protein in yellow perch pituitaries and also recognizes a purified sea bream GH preparation. However, it did not recognize any bands on Western blots of brook trout pituitaries. The antibody was used to measure the levels of GH immunoreactive protein during the year. The highest levels of GH were observed during May and the lowest levels in January. In yellow perch that were treated with estradiol-beta (E2) in the diet for 46 days, there were significantly elevated levels of GH compared to controls and these results are consistent with past published reports on the stimulatory effects of E2 on growth in yellow perch. The antibody that was generated in this study can be used to measure GH in experiments aimed at understanding the factors that affect yellow perch growth. This should facilitate the optimization of these factors for aquaculture production.
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