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Expression，Purification and Bioinformatics Analysis of β-hexosaminidase of Dermatophagoides farinae

MO Li-hua1，LIU Yu-lin2，YANG Li-tao3，FAN Xiao-qin3，LIU Zhi-gang1，YANG Ping-chang 1 *   

  1. 1 Institute of Allergy and Immunology，Shenzhen University，Shenzhen 518060，China；2 Department of Immunology，Medical College of Nanchang University，Nanchang 330006，China；3 Institute of Otolaryngology，Shenzhen University，Shenzhen 518020，China
  • Online:2015-04-30 Published:2015-05-04

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Abstract:

The DNA fragment encoding β-hexosaminidase was synthesized， and cloned into pET-28a vector. The constructed plasmid pMD18-T-β-hexosaminidase was transformed into E. coli Top10 and followed by expression of the protein induced by IPTG. SDS-PAGE result showed that the relative molecular mass of the recombinant protein was about Mr 55 000. The full length of β-hexosaminidase gene was 1 410 bp. Bioinformatics analysis revealed that β-hexosaminidase was composed with 469 amino acid residues with a calculated molecular weight of Mr 55 000， and its secondary structure was composed of strand（14.71%）， helix（30.70%）， and loop（54.58%）. β-hexosaminidase was a hydrophilic protein without signal peptide， and located in the extracellular space.

Key words: Dermatophagoides farinae；&beta, -hexosaminidase；Prokaryotic expression