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Molecular Cloning and Expression of Grass Carp MyoD in Yeast Pichia pastoris
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  • Molecular Cloning and Expression of Grass Carp MyoD in Yeast Pichia pastoris
  • Molecular Cloning and Expression of Grass Carp MyoD in Yeast Pichia pastoris
저자명
Wang. Lixin,Bai. Junjie,Luo. Jianren,Chen. Hong,Ye. Xing,Jian. Qing,Lao. Haihua
간행물명
Journal of biochemistry and molecular biology
권/호정보
2007년|40권 1호|pp.22-28 (7 pages)
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생화학분자생물학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

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.