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

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...

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Main Authors: Mohd Rashid, Noor Fazielawanie, Siraj, Siti Shapor, Harmin, Sharr Azni, Md Yasin, Ina Salwany
Format: Article
Language:English
Published: Springer 2013
Online Access:http://psasir.upm.edu.my/id/eprint/29231/1/29231.pdf
http://psasir.upm.edu.my/id/eprint/29231/
http://link.springer.com/article/10.1007%2Fs10695-012-9690-5
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spelling my.upm.eprints.292312016-04-25T07:04:02Z http://psasir.upm.edu.my/id/eprint/29231/ Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin Mohd Rashid, Noor Fazielawanie Siraj, Siti Shapor Harmin, Sharr Azni Md Yasin, Ina Salwany 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. Springer 2013 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/29231/1/29231.pdf 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 http://link.springer.com/article/10.1007%2Fs10695-012-9690-5 10.1007/s10695-012-9690-5
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description 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.
format Article
author Mohd Rashid, Noor Fazielawanie
Siraj, Siti Shapor
Harmin, Sharr Azni
Md Yasin, Ina Salwany
spellingShingle Mohd Rashid, Noor Fazielawanie
Siraj, Siti Shapor
Harmin, Sharr Azni
Md Yasin, Ina Salwany
Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin
author_facet Mohd Rashid, Noor Fazielawanie
Siraj, Siti Shapor
Harmin, Sharr Azni
Md Yasin, Ina Salwany
author_sort Mohd Rashid, Noor Fazielawanie
title Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin
title_short Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin
title_full Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin
title_fullStr Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin
title_full_unstemmed Isolation and partial characterization of Asian sea bass (Lates calcarifer) vitellogenin
title_sort isolation and partial characterization of asian sea bass (lates calcarifer) vitellogenin
publisher Springer
publishDate 2013
url http://psasir.upm.edu.my/id/eprint/29231/1/29231.pdf
http://psasir.upm.edu.my/id/eprint/29231/
http://link.springer.com/article/10.1007%2Fs10695-012-9690-5
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score 13.222552