Purification of beluga (Huso huso) vitellogenin using selective precipitation (EDTA-Mg+2) and excision from reducing gel preparative

Document Type : Research Paper

Authors

1 MSc graduate, Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran

2 Professor, Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran

3 Associate Prof. Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran

4 Assistant Prof. Industrial and Environmental Biotechnology Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), 14965-161, Tehran, Iran

Abstract

Vitellogenin (Vtg) is a glycophospholipoprotein precursor of egg yolk which circulating levels of this protein in male and juvenile fish provide a sensitive indication of exposure to endocrine disruptor chemicals. The objectives of the present study were identification and purification of beluga (Huso huso) Vtg using Vtg selective precipitation (EDTA-Mg+2) and excision of Vtg band from reducing gel preparative. In this study, required plasma samples were collected from vitellogenic beluga and Vtg was identified using the specific characteristics of this protein (range of molecular weight, specificity for female, estrogen inducibility, precipitation in the presence of EDTA as well as specific reactivity of putative beluga Vtg with antibodies to beluga Vtg). Vtg highly purified by combination of Vtg selective precipitation and gel preparative. With SDS-PAGE under reducing conditions, beluga Vtg appeared as a major band of ~200 KDa. These data suggest that vitellogenin selective precipitation followed by gel preparative is an efficient procedure for the characterization and purification of beluga Vtg.

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