High Expression and Preliminary Purification of Human β-Defensin-2 Fusion Protein

Abstract:

This study was undertaken to achieve high expression and preliminary purification of human β-defensin-2 fusion protein to lay a solid foundation for production of human β-defensin-2 using genetic engineering. A prokaryotic expression vector for human β-defensin-2 fusion protein was generated using in vitro gene synthesis before transformation into BL21 (l DE3) plysS TrX-B host bacteria. High expression of TrX-A-HBD-2 fusion protein was induced with IPTG in the bacteria exposed to various expression conditions. The fusion protein then underwent preliminary purification. The protein of interest was released from the genetically engineered bacteria after freezing and thawing. The expression of the target protein accounted for 16.12% of the total bacterial proteins. Fractional precipitation with saturated ammonium sulfate and metal chelate affinity chromatography yielded human β-defensin-2 peptide fusion protein, with a relative purity of 80.53%.Human β-defensin-2 fusion protein could be highly expressed in a soluble form, with a relatively high purity

• Periodical: Advanced Materials Research (Volumes 781-784)
• Pages: 1076-1079
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.781-784.1076
• Online since: September 2013
• Authors: Hong Tao Wei, Zhong Wen Lv, Xue Mei Han, Guo Li Zhang
• Keywords: Fusion Protein, Gene Recombination, Human β-defensin-2

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