鸭β-防御素16的分离、鉴定及其抗病毒机制

doi:10.3864/j.issn.0578-1752.2012.18.020

Abstract

Abstract: 【Objective】The objective of the present study was to clone, express, and characterize the antimicrobial activity of duck avian beta-defensin (AvBD16). AvBD16 and TLR-7 dynamic changes were evaluated in different duck organizations after duck hepatitis virus infection. 【Method】The specific cDNA was obtained by PCR from bone marrow of ducks. The cDNA of duck AvBD16 was cloned into pGEX-6p-1 vector to construct recombinant plasmid, which were transformed into E. coli BL21 and the bacteria were induced with IPTG. Furthermore, the recombinant protein was purified. Peptides were synthesized according to the gene sequence. The antimicrobial activity of both recombinant and synthetic AvBDs was investigated in vitro. In addition, the mRNA expressions of AvBD16 and TLR-7 in tissues after duck hepatitis virus (DHV) infection were examined.【Result】Duck AvBD16 consisted of 155 bp encoding 50 amino acids and shared an amino acid homology (62%) with chicken AvBD3. Both recombinant and synthetic duck AvBD16 showed similar antibacterial activities. In high salt ions conditions, antibacterial activity of duck AvBD16 protein was decreased. In addition, the hemolysis activity of the peptide was extremely low. The duck AvBD16 exhibited significant antiviral activity against DHV in vitro. The mRNA expression of AvBD16 and TLR-7 in most tissues, including immune organs and liver, was upregulated or induced in response to DHV infection at different time points.【Conclusion】The duck AvBD16 gene from duck was successfully cloned, expressed in E. coli, and recombinant and synthetic AvBD16 protein showed antimicrobial activity and has no hemolytic properties. Duck AvBD16 exhibits significant antiviral activity against DHV in vitro. This result suggests that the antiviral activity against DHV of the AvBD16 is partially via a TLR7-dependent mechanism.

Key words: duck AvBD16, antibacterial activity, antiviral activity, signaling transduction

ZHANG Ke-Xin, ZHANG Ming-Yue, XIN Sheng-Nan, HAN Zong-Xi, SHAO Yu-Hao, LIU Sheng-Wang, MA De-Ying. Isolation, Characterization and Mechanism of Anti-viral Activity of Duck Avian Beta-defensin 16[J].Scientia Agricultura Sinica, 2012, 45(18): 3873-3882.

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