Bo Tian

Publications (2)4.21 Total impact

• Article: High-level expression and secondary structure analysis of the bovine mature prion protein

  Dawei Wang, Huaiyi Yang, Zihe Rao, Bo Tian
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  ABSTRACT: By using the recombinant DNA technology, the gene of the bovine mature prion protein (bPrPCL) has been cloned into pET30a and the resulting plasmid has been expressed inE.coli BL21(DE3). After solubilizing in 8 mol/L urea, the expression product was purified by cation ion exchange chromatography. The purified product was refolded by dilution and the recovery was about 15%. Analysis of mass spectrum, circular dichroism (CD) spectrum and Fourier transform infrared (FTIR) spectrum demonstrate that the molecular weight of the bPrPCL is 23 630 u, the bPrPCL has a high α-helix content (36.1%) and low β-sheet content (11.9%). KeywordsbPrPCL-expression-CD-FTIR-secondary structure
  Chinese Science Bulletin 04/2012; 45(14):1312-1315. · 1.32 Impact Factor
• Article: Purification of recombinant bovine normal prion protein PrP(104-242) by HPHIC.

  Chaozhan Wang, Xindu Geng, Dawei Wang, Bo Tian
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  ABSTRACT: Purification of the prion protein (PrP) is a major concern for biological or biophysical analysis as are the structural specificities of this protein in relation to infectivity. A simple and efficient method for purification of recombinant bovine normal prion protein containing residues 104-242, PrP(104-242) expressed in Escherichia coli by high performance hydrophobic interaction chromatography (HPHIC) was presented in this work. The solution containing denatured and reduced protein in 8.0 mol/L urea extracted from the inclusion body was directly injected into the HPHIC column, aggregates were prevented by the interaction between the denatured PrP(104-242) molecules and the stationary phase during the chromatographic process, the soluble form of PrP(104-242) in aqueous solution was obtained after desorbed from the column. Several factors, including pH value, types of stationary phase and salt, and gradient mode, influencing the purification results were investigated. Optimal conditions were obtained for the purification of PrP(104-242) by HPHIC. This procedure yield PrP(104-242) of a purity of 96% with a recovery of 87%, respectively, for a single step purification of 40 min.
  Journal of Chromatography B 08/2004; 806(2):185-90. · 2.89 Impact Factor