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

文献类型： 外文期刊

第一作者： Wang, Lixin

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

作者机构：

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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