Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (5): 999-1009.doi: 10.3864/j.issn.0578-1752.2012.05.022

• VETERINARY SCIENCE • Previous Articles     Next Articles

Isolation, Characterization, and Expression of Goose Avian β-Defensin 10

 ZHOU  Cai-Yuan, ZHANG  Ming-Yue, HAN  Zong-Xi, SHAO  Yu-Hao, LIU  Sheng-Wang, MA  De-Ying   

  1. 1.东北农业大学动物营养研究所，哈尔滨 150030
    2.中国农业科学院哈尔滨兽医研究所/兽医生物技术国家重点实验室禽传染病研究室，哈尔滨 150001
  • Received:2011-02-28 Online:2012-03-01 Published:2011-07-20

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Abstract: 【Objective】 The objective of this study was to clone avian β-defensin 10 (AvBD10) gene from goose tissues and to characterize the antimicrobial activity of the recombinant GST-AvBD10 fusion protein. 【Method】 The coding sequences of goose AvBD10 was obtained from kidney of goose by RT-PCR, phylogenetic relationships of the goose AvBD10 with those of other avian species and some mammalian were analyzed. In addition, tissue distribution of the gene was detected by real time PCR. AvBD gene coding sequence was inserted into the pGEX-6p-1 vector. The constructs were transformed into competent Escherichia coli BL21 (DE3) cells. Expression of the fusion proteins was induced with isopropyl β-D-1-thiogalactopyranoside (IPTG) and the proteins were purified. Furthermore, bioactivities of the recombinant protein were analyzed in vitro. 【Result】 The full length cDNA of goose AvBD10 consisted of 207 nucleotide acids, encoding 68 amino acid residues. Phylogenetic analysis demonstrated that the goose AvBD10 shared the highest amino acid homology with duck AvBD10 (92.7%). The recombinant protein exhibited high antimicrobial activity against 12 bacterial strains investigated. The antimicrobial activity was decreased in high salt concentration and exhibited no hemolytic properties. 【Conclusion】 The goose AvBD10 gene from goose was successfully cloned and expressed in E. coli. The purified recombinant protein showed antimicrobial activity and no hemolytic properties.

Key words: goose AvBD10, clone, phylogenetic analysis, tissue distribution, antimicrobial activity

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