Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Effect of Recombinant Olive Flounder Stanniocalcin on Serum Calcium Levels

혈청 칼슘 농도에 미치는 넙치 유전자 재조합 스타니오칼신의 효과

  • Shin, Ji-Hye (Department of Marine Molecular Biotechnology, Faculty of Marine Bioscience and Technology, Gangneung-Wonju National University) ;
  • Jung, Yu-Jung (Regeron, Inc.) ;
  • Han, Yoon-Hee (Regeron, Inc.) ;
  • Lee, Kyun-Young (Regeron, Inc.) ;
  • Lee, Kyung-Mi (Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, University of Tokyo) ;
  • Kaneko, Toyoji (Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, University of Tokyo) ;
  • Sohn, Young-Chang (Department of Marine Molecular Biotechnology, Faculty of Marine Bioscience and Technology, Gangneung-Wonju National University)
  • Received : 2009.12.02
  • Accepted : 2010.08.13
  • Published : 2010.08.31
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Abstract

Stanniocalcin 1 (STC1) is a glycoprotein hormone that is important in the maintenance of calcium and phosphate homeostasis in both fish and mammals. STC1 and its paralog STC2 are expressed in multiple tissues in fishes, although the physiological roles of piscine STCs are still unclear compared with those of mammals. In this study, we cloned olive flounder STC1 (ofSTC1) and ofSTC2 cDNAs into pET28a vector and used E. coli Rosetta (DE3) as the host strain for protein expression. Expression experiments were carried out using isopropyl-$\beta$-D-thiogalactoside (IPTG) and nickel affinity chromatography. We could identify the recombinant proteins as single 29.5 kDa (ofSTC1) and 33.2 kDa (ofSTC2) bands in the insoluble fraction on sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). These results indicate that ofSTC1 and ofSTC2 were expressed as insoluble proteins in E. coli. Furthermore, the injection of ofSTC1 protein into juvenile tilapia resulted in a decrease of the serum calcium level. These results suggest that the purified fish STC1 and STC2 proteins may be used to elucidate the physiological role of STCs in fishes.

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References

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