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Molecular cloning and expression of grass carp MyoD in yeast Pichia pastoris

文献类型: 外文期刊

作者: Wang, Lixin 1 ; Bai, Junjie 2 ; Luo, Jianren 2 ; Chen, Hong 2 ; Ye, Xing 2 ; Jian, Qing 3 ; Lao, Haihua;

作者机构: 1.CAFS, Pearl River Fisheries Res Inst, Key Lab Trop & Subtrop Fish Breeding & Cultivat, Guangzhou 510380, Peoples R China

2.CAFS, Pearl River Fisheries Res Inst, Key Lab Trop & Subtrop Fish Breeding & Cultivat, Guangzhou 510380, Peoples R China; NW A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Shaanxi, Peoples R China

3.CAFS, Pearl River Fisheries Res Inst, Key Lab Trop & Subtrop Fish Breeding & Cultivat, Gua

关键词: expression;grass carp;MyoD gene;molecular cloning;Pichia pastoris

期刊名称:JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY ( 影响因子:2.021; 五年影响因子:2.1 )

ISSN: 1225-8687

年卷期: 2007 年 40 卷 1 期

页码:

收录情况: SCI

摘要: 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 (1-84(th) amino acids) and HLH domain (98-142(th) 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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