Shuang Feng

Publications (2)1.38 Total impact

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  Available from: Tianbo Li

  Article: [Expression of a human myofibrillogenesis regulator 1 gene in E. coli and its immunogenicity].

  Tian-bo Li, Yang Hu, Shuang Feng, Zeng-yan Zuo, Yi-guang Wang
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  ABSTRACT: To study the expression of human myofibrillogenesis regulator 1 (MR-1) gene in E. coli and obtain the MR-1 protein and its antibody for further investigation of its biological function. Expression vectors pGEX-5X-1, pET30a (+), and pET24a (+), as well as host strain E. coli BL21 (DE3) and BL21-CodonPlus (DE3) -RIL were used for expression of MR-1. MR-1 N-terminal with GST or T7-tag or C-terminal with His-tag, separately, or N terminal with T7-tag and C terminal with His-tag, simultaneously, were fused in plasmids pGEX-5X-1, pET30a (+) , and pET24a (+). The expressed MR-1-T protein, separated and purified by preparative SDS-PAGE, was applied to immunize the rabbits. The titer of the antibody was assayed by ELISA and its immunogenicity was tested by Western blot with pcDNA3/MR-1 transfected human breast cancer cell MCF7. The MR-1 protein was successfully expressed as inclusion body by fusing its N-terminal with T7-tag in E. coli BL21-CodonPlus (DE3) -RIL. MR-1 protein was purified by electro-elution from SDS-PAGE gel. Using this purified protein, polyclonal antibody in rabbit against MR-1 was essentially generated. ELISA and Western blot showed the titer of this antibody was about 1:10(5) with high immunogenicity. The N-terminal fusion tag is the most important mechanism for MR-1 expression. The polyclonal antibody of the generated MR-1 protein in E. coli may be applied for its further biological function studies.
  Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae 03/2005; 27(1):42-7.
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  Available from: Tianbo Li

  Article: Characterization of MR-1, a novel myofibrillogenesis regulator in human muscle.

  Tian-Bo Li, Xiu-Hua Liu, Shuang Feng, Yang Hu, Wei-Xi Yang, Yue Han, Yi-Guang Wang, Li-Min Gong
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  ABSTRACT: The actin-myosin contractile apparatus consists of several thick filament and thin filament proteins. Specific regulatory mechanisms are involved in this highly ordered process. In this paper, we reported the identification and characterization of a novel myofibrillogenesis regulator, MR-1. The MR-1 gene was cloned from human skeletal muscle cDNA library by using a strategy that involves EST data base searching, PCR and RACE. The MR-1 gene is located on human chromosome 2q35 and encodes a 142 aa protein. Northern blot revealed that the mRNA level of MR-1 was highest in the skeletal muscle and certain level of MR-1 expression was also observed in heart, liver and kidney. Immunohistochemical assay confirmed that the MR-1 protein existed in human myocardial myofibrils. It was found by yeast two-hybrid screening and confirmed by in vitro binding assay that MR-1 could interact with sarcomeric proteins, such as myosin regulatory light chain, myomesin 1 and beta-enolase. These studies suggested that MR-1 might play a regulatory role in the muscle cell and it was worth investigating further.
  Acta Biochimica et Biophysica Sinica 07/2004; 36(6):412-8. · 1.38 Impact Factor