GP5Δ84-119稳定表达对猪繁殖与呼吸综合征病毒复制的影响

doi:10.11843/j.issn.0366-6964.2016.02.014

Abstract

Abstract:

This experiment was conducted to study the role of the second extracellular domain of glycoprotein 5，one major structural protein of porcine reproductive and respiratory syndrome virus (PRRSV)，on PRRSV replication and approach to its mechanism in Marc-145 cells.A recombinant plasmid (named as pPB-^GP5Δ84-119) expressing truncated GP5 was constructed using PiggyBac Transposon System Vectors and transfected to Marc-145 cells.Marc-145 cell line stably expressing GP5^Δ84-119 was obtained by puromycin resistance screening and triple subcloning and the selected cells proliferation was detected using cck-8 kit.The effect of GP5^Δ84-119 stable expression on PRRSV replication were determined thought detection of virus gene copy number in the infected cells and virus titers in the supernatant of infected cells.What’s more，the mRNA and protein expression levels of IFN-α，IFN-β，IFN-γ in the cells before and after PRRSV infection were also analyzed using Real-time PCR and ELISA.The results of RT-PCR，Western blot and IFA confirmed that GP5^Δ84-119 was stablely expressed in Marc-145 cells and the cells were named as Marc-145-GP5^Δ84-119.The result of cell proliferation assay confirmed that the expression of GP5^Δ84-119 did not affect the proliferation of Marc-145 cells.It was found that GP5^Δ84-119 expression inhibited the replication of highly pathogenic PRRSV in Marc-145 cells through upregulating IFN level，especially IFN-β.These findings suggest that the second extracellular domain of GP5 plays an important role in PRRSV replication.

CLC Number:

S852.659.6

WANG Xiao-hong，SONG Lin-lin，LI Liang-liang，YUAN Chuan-qi，JIANG Bo，ZHOU En-min，MU Yang. The Effect of GP5^Δ84-119 Stable Expression on the Replication of Porcine Reproductive and Respiratory Syndrome Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(2): 315-324.

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The Effect of GP5^Δ84-119 Stable Expression on the Replication of Porcine Reproductive and Respiratory Syndrome Virus

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The Effect of GP5Δ84-119 Stable Expression on the Replication of Porcine Reproductive and Respiratory Syndrome Virus

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The Effect of GP5^Δ84-119 Stable Expression on the Replication of Porcine Reproductive and Respiratory Syndrome Virus