CRCU

Chemical Research in Chinese Universities ›› 2006, Vol. 22 ›› Issue (2): 157-161.

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Expression and Characterization of Human Heart Type Fatty Acid Binding Protein in Pichia Pastoris

HOU Wei1, TAN Yan2, XU Shu-fen2, YANG Xiao-hong1, ZHANG Shu-hua3, LIU Ling-li2, CHE Yuan-yuan2, LIU Li-hua2   

  1. 1. Pharmacy College of Jilin University;
    2. The First Hospital of Jilin University, Changchun 130021, P.R.China;
    3. Changchun University of Science and Technology, Changchun 130021, P.R.China
  • Received:2005-12-31 Online:2006-04-24 Published:2011-08-06
  • Supported by:

    Supported by the Social Development Project of Science and Technology Department of Jilin Province, China(No.20030450).

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Abstract: H-FABP is regarded as a tissue-specific protein existing only in myocardial cells.It is released from the cardiac tissue and gets into the plasma when a heart attack occurs; the myocardial infarction is a good case in point.As a result, the detection of H-FABP will be an early and important biomarker for the disease concerned.The objective of the study is to prepare the recombinant H-FABP by aeukaryotic expression system, pichia, to produce the protein mimicking natural H-FABP, as an immunogen for the production of the specific antibody.A gene fragment encoding H-FABP was cloned in the expressing vector pPICZα, after sequencing.The recombinant plasmid was transformed into the competent cells of the X-33 strain by means of electroporation.The expression of the target peptide indueed by methanol was screened by means of Western blotting, with the available MAb(Clone 6B6).Highly expressive engineer strains were obtained.The production of recombinant H-FABP under induction was about 0.7 g/L, with an Mr of 14.5 kDa and recognized by a commercially available MAb (Clone 6B6).The recombinant vector was successfully constructed.Following this, H-FABP was expressed in X-33, and it would become the source of the preparation of specific antibodies, to develop diagnostic kits.

Key words: Human heart type fatty acid binding protein, Expression and purification, Pichia