猪肿瘤坏死因子受体相关因子6的克隆、表达及对伪狂犬病病毒的抑制效应

doi:10.11843/j.issn.0366-6964.2017.02.012

Abstract

Abstract:

This study aimed to explore the antiviral effect and immunomodulatory functions of porcine tumor necrosis factor receptor-associated factor 6 (TRAF6), an important adaptor molecule in innate immune signaling pathway. Firstly, porcine TRAF6 gene was cloned by RT-PCR from porcine peripheral blood mononuclear cells and gene sequence was analyzed by sequence alignment. Secondly, stably expressing porcine TRAF6 cell line was established by Piggybac transposon system. And finally, we detected the impacts of TRAF6 on Pseudorabies virus (PRV) replication and porcine IFN-β promoter activity at the cellular level. The results showed that porcine TRAF6 gene was 1 623 bp. It was connected to Piggybac transposon (PB) plasmid and successfully constructed the eukaryotic expression vector PB-TRAF6, and then the recombinant plasmid was transfected into PK-15 cells to establish stably-expressed cell line by drug treatment and fluorescence screening. The high-expression of TRAF6 was confirmed by real time PCR and Western blot in the cell line. In vitro antiviral tests showed that the replication of PRV could be significantly inhibited in the cell line. Dual luciferase reporter system showed that porcine TRAF6 significantly enhanced poly(I:C)-mediated type Ⅰ interferon responses. These results indicated that porcine TRAF6 possesses significant anti-PRV activity and immunomodulatory functions, which will provide the basis for further research on immunology functions of TRAF6.

CLC Number:

S858.285.3

LI Yong-tao, LI Hong-jie, WANG Xiao-xue, JIA Guang-min, CHEN Lu, ZHAO Jun, WANG Chuan-qing, LIU Hong-ying. Cloning, Expression and Antiviral Effect against Pseudorabies Virus of Porcine Tumor Necrosis Factor Receptor-associated Factor 6[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 289-296.

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Cloning, Expression and Antiviral Effect against Pseudorabies Virus of Porcine Tumor Necrosis Factor Receptor-associated Factor 6

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