Studied On Purification,Characterization Identification And Seasonal Content Variation Of Vitellongin From Dolly Varden Charr,Salvelinus Malma

The vitellogenin content of seasonal variation in the blood of Salvelinus malma was determined by the use of the immunological methods in this experiment.Through the researches on the change of the content,the physicochemical properties of vitellogenin and the relationship between the vitellogenin quantity and sexual maturity can be better understood.These conclusions could provide theoretical basis for the research on the occurrence and synthetic pathway of vitellogenin of salmonidae fish.In this experiment,3 age Salvelinus malma was used to produce vitellogenin by the intraperitoneal injection of estradiol.The vitellogenin was isolated and purified with Sephacryl S-300 and then its physicochemical properties were studied.The result shows that Sephacryl S-300 could isolate and purify the vitellogenin in blood very well.The purified vitellogenin was conducted Native-PAGE and SDS-PAGE,and the electrophoresis results showed that the molecular weight was about 590 kDa and subunit was about200 kDa.The vitellogenin was used as antigen to immune rabbit,and then antiserum was prepared and the specificity of the antiserum was detected.The indirect competitive enzyme linked immunosorbent assay was established to detect the vitellogenin content in the blood of Salvelinus malma was established because of high specificity of the antiserum.The optimum concentration of antiserum and antigen coating were 1:32000 and 130.7 ng/mL.The linear detection ranged from 32.6ng/mL to 2094.5 ng/mL,with mean intra-and inter-assay coefficients of variation were 4.612% and 4.337%,respectively.Seasonal variation of vitellogenin content in the blood of Salvelinus malma was determined by the use of ELISA.The result showed the vitellogenin content changed in the periodical,and the content of vitellogenin raised to a very high level in June before hatching then decreased gradually to a very low level in December after hatching and then followed with the content increased again.This experiment shows that vitellogenin exists only in the female Salvelinus malma.Vitellogenin is a precursor of the vitellin,and the vitellin is very important in early embryos and larvae of development.Therefore,vitellogenin is significant for fish breeding and nutrition as well.The vitellogenin level in the blood changed constantly.The conclusion of this experiment provides the experimental basis and theoretical basis for fully understanding of the propagation characteristics of Salvelinus malma,and it also makes great significance for the protection,development and utilization of Salvelinus malma.