BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Molecular Cloning and Expression of Grass Carp MyoD in Yeast Pichia pastoris

  • Wang, Lixin (Pearl River Fisheries Research Institute, CAFS. Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation of CAFS) ;
  • Bai, Junjie (Pearl River Fisheries Research Institute, CAFS. Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation of CAFS) ;
  • Luo, Jianren (Pearl River Fisheries Research Institute, CAFS. Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation of CAFS) ;
  • Chen, Hong (College of Animal Science and Technology, Northwest Science and Technology University of Agriculture and Forestry) ;
  • Ye, Xing (Pearl River Fisheries Research Institute, CAFS. Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation of CAFS) ;
  • Jian, Qing (Pearl River Fisheries Research Institute, CAFS. Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation of CAFS) ;
  • Lao, Haihua (Pearl River Fisheries Research Institute, CAFS. Key Lab. of Tropical & Subtropical Fish Breeding & Cultivation of CAFS)
  • Published : 2007.01.31
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Abstract

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

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References

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