Journal of Microbiology and Biotechnology

  • 제23권3호
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  • Pages.322-328
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  • 2013
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Expression and Purification of Human Farnesoid X Receptor-Ligand Binding Domain as Soluble Form Using a Dual Cistronic Expression Vector

  • Kang, Hyun (Department of Biotechnology, College of Biomedical and Health Science, Konkuk University) ;
  • Ye, Micheal B. (Department of Applied Biochemistry, College of Biomedical and Health Science, Konkuk University) ;
  • Bahk, Young Yil (Department of Biotechnology, College of Biomedical and Health Science, Konkuk University)
  • 투고 : 2012.12.05
  • 심사 : 2012.12.21
  • 발행 : 2013.03.28
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초록

In this study, we show the expression and purification of the human recombinant farnesoid X receptor (FXR)- ligand binding domain (LBD) protein in E. coli using a double cistronic vector, pACYCDuet-1, as a soluble form. We describe here the expression and characterization of a biologically active $FXR-LBD_{(248-476)}$. When expressed in the influence of bacterial promoters ($P_{T7}$ and $P_{Tac}$) of the single cistronic expression vectors, the human recombinant $FXR-LBD_{(248-476)}$ was found to be totally insoluble. However, by using a double cistronic expression vector, we were able to obtain the human recombinant $FXR-LBD_{(248-476)}$ in a soluble form. To allow for biological activities, we have subcloned into the pACYCDuet-1 vector, expressed in E. coli cells at some optimized conditions, and purified and characterized the human recombinant active $FXR-LBD_{(248-476)}$ proteins using the fluorescence polarization assay. This suggests that the expression of FXR-LBD in a double cistronic vector improves its solubility and probably assists its correct folding for the biologically active form of the proteins. We suggest that this may represent a new approach to high expression of other nuclear receptors and may be useful as well for other classes of heterodimeric protein partners.

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