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Vitellogenin isolation, purification and antigenic cross-reactivity in three teleost species

Citation

Watts, M and Pankhurst, NW and Pryce, A and Sun, Biao, Vitellogenin isolation, purification and antigenic cross-reactivity in three teleost species, Comparative Biochemistry and Physiology. Part B: Biochemistry and Molecular Biology, 134, (3) pp. 467-476. ISSN 1096-4959 (2003) [Refereed Article]

DOI: doi:10.1016/S1096-4959(02)00288-9

Abstract

Vitellogenin (Vtg) was isolated from male greenback flounder (Rhombosolea tapirina), rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) plasma, following induction by estradiol (E 2) inoculation. The molecular weight of each native molecule, as determined by gel filtration, was 540, 383 and 557 kDa, respectively. With sodium dodecyl sulphate polyacrylamide gel electrophoresis under reducing conditions, Atlantic salmon and greenback flounder Vtg appeared as three major bands (∼159, 117, 86 kDa and 155, 104, 79 kDa, respectively), whereas rainbow trout Vtg appeared as one major band (∼154 kDa). Several minor bands were also present in each Vtg isolate. Polyclonal antisera, produced against only the highest molecular weight band from each species following excision from reducing gels, were reactive with all major bands in Western blots. In competition ELISA, parallel binding slopes were demonstrated between purified Vtg and plasma from vitellogenic females of the same species, but there was no reaction with plasma from untreated males. These antisera were highly species-specific and little cross-reactivity was noted, even between the two salmonid species. These data suggest that excision of bands from gels is a simple procedure for the preparation of species-specific antisera, and confirm that cross-species assays give highly variable results. © 2002 Elsevier Science Inc. All rights reserved.

Item Details

Item Type:Refereed Article
Research Division:Agricultural and Veterinary Sciences
Research Group:Fisheries Sciences
Research Field:Aquaculture
Objective Division:Animal Production and Animal Primary Products
Objective Group:Fisheries - Aquaculture
Objective Field:Fisheries - Aquaculture not elsewhere classified
Author:Watts, M (Dr Marianne Watts)
Author:Pankhurst, NW (Professor Ned Pankhurst)
Author:Sun, Biao (Dr Biao Sun)
ID Code:49431
Year Published:2003
Web of Science® Times Cited:28
Deposited By:Health Sciences A
Deposited On:2007-12-11
Last Modified:2011-09-30
Downloads:0

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