Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (8): 108-112.

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Cloning,Sequence Analysis and Prokaryotic Expression of Musca domestica Allergen Tropomyosin Gene

Wei Chuanchuan, Xiu Jiangfan, Wang Yu, Guo Guo, Peng Chuanlin, Wu Qinyi, Wu Jianwei   

  1. Basic Medical College, Guiyang Medical College, Guiyang 550004
  • Revised:2014-01-08 Online:2014-08-15 Published:2014-08-01
  • Contact: 通讯作者: 吴建伟,男,教授,博士生导师,研究方向:昆虫分子免疫学研究;E-mail:wjw@gmc.edu.cn
  • Supported by:

    国家科技支撑计划(2011BAC06B12);国家自然科学基金项目(81360254);贵州省科学技术基金项目(黔科合J字[2012]2038号)

Abstract:

It was to probe into the homologous cloning of the Musca domestica allergen Tropomyosin gene, and then construct the prokaryotic expression vector which was expressed in Escherichia coli. Screening and isolation of Musca domestica Tropomyosin gene from Musca domestica cDNA library was carried out. The gene and encoded protein sequence of Tropomyosin was analyzed by bioinformatics methods , including general physical and chemical properties, signal peptide, secondary structure, tertiary structure, epitope and subcellular localization. Then plasmid pEASY-E1-Tropomysin were transformed into Escherichia coli BL21(DE3) competent cells for expression. The open reading frames of the Tropomyosin gene was 828 bp that encded a putative protein with 275 amino acids. The protein, with predicted molecular weight 31.6 kD and pI of 4.65, has the conserved Tropomyosin domain so belongs to Tropomyosin family. The result showed that the recombinant prokaryotic expression vector pEASY-E1-Tropomysin was successfully constructed and fusion protein was expressed in E. coli.

Key words: Musca domestic, Tropomyosin, Cloning, Bioinformatics, Allergen