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Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin


Mohd Rashid, Noor Fazielawanie and Siraj, Siti Shapor and Harmin, Sharr Azni and Md Yasin, Ina Salwany (2013) Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin. Fish Physiology and Biochemistry, 39 (2). pp. 191-200. ISSN 0920-1742; ESSN: 1573-5168


A study was conducted to isolate, partial characterize Asian sea bass (Lates calcarifer) vitellogenin (vtg). Two-year-old juvenile L. calcarifer (n = 10) were given three intraperitoneal injections of 17-β estradiol (E2) at a dose of 2 mg/kg body weight to induce vitellogenesis. Blood was collected 3 days after the last injection, and plasma was purified through gel filtration chromatography. A broad single symmetrical peak consisting of vtg molecule was produced. Protein concentration was 0.059 mg/ml as determined by Bradfrod assay using bovine serum albumin as a standard. The protein appeared as one circulating form in Native PAGE considering the dimeric form of putative vtg with molecular weight of 545 kDa. In SDS-PAGE under reducing conditions, two major bands appeared at 232.86 and 118.80 kDa and minor bands at 100.60, 85.80 and 39.92 kDa, respectively. The purified vtg was used to generate a polyclonal antibody, and the specificity of antibody was assessed by Western blot analysis. Two major bands were immunoreacted, but no cross-reactivity was observed with plasma from non-induced males. The protein was characterized as phosphoglycolipoprotein as it positively stained for the presence of lipid, phosphorus and carbohydrate using Sudan Black B, methyl green and periodic acid/Schiff reagent solution, respectively. The amino acid composition was analyzed by high sensitivity amino acid analysis that showed high percentage of non-polar amino acids (~48 %). The results suggest the potential utilization of vtg as a basis tool to further study about reproductive physiology of this important economical species.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Agriculture
DOI Number: https://doi.org/10.1007/s10695-012-9690-5
Publisher: Springer
Keywords: Amino acid analysis; Chromatography; Lates calcarifer; Phosphoglycolipoprotein; Vitellogenin; Western blot
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 25 Apr 2016 07:04
Last Modified: 25 Apr 2016 07:04
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1007/s10695-012-9690-5
URI: http://psasir.upm.edu.my/id/eprint/29231
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