Influence of Nsp9 Between Low Virulent and Virulent Strains to Replication of PRRSV in Marc-145 Cells

doi:10.7668/hbnxb.2017.01.003

Abstract

Abstract: In order to verify the difference of gene Nsp9 between virulent strain and low virulent strain whether could enchance the replication of PRRSV, plasmids pIRes2-EGFP- Nsp9 -XH-GD and pIRes2-EGFP-Nsp9-CH-1R containing whole Nsp9 genome were transfected into Marc-145 cells,qPCR and Western Blot were conducted to evaluate its titer and expression after PRRSV was inoculated,the results showed that the level of N protein in the cells that were transfected with low virulent strain CH-1R Nsp9 gene was higher than the control group-virulent strain XH-GD in the mRNA level. Meanwhile, on the protein level, it was the same as the mRNA level, and virus titer was also higher than virulent strain. It demonstrated that Nsp9 gene of low virulent strain CH-1R in the Marc-145 cells was more beneficial to the replication of PRRSV than that of virulent strain XH-GD.

Key words: PRRSV, Nsp9, Replication, Viraltiter

CLC Number:

ZHAO Mengmeng, FENG Lili, FENG Songlin, MA Hongfang, ZHANG Erqin, WANG Wenjia, XING Xing, ZHANG Ya, FENG Jiaping, ZHANG Guihong. Influence of Nsp9 Between Low Virulent and Virulent Strains to Replication of PRRSV in Marc-145 Cells[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(1): 15-20. doi: 10.7668/hbnxb.2017.01.003.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2017.01.003

http://www.hbnxb.net/EN/Y2017/V32/I1/15

References

[1] Andreyev V G,Wesley R D,Mengeling W L,et al.Genetic variation and phylogenetic relationships of 22Porcine reproductive and respiratory syndrome virus(PRRSV) field strains based on sequence analysis of open reading frame 5[J].Archives of Virology,1997,142(5):993-1001.
[2] Benfield D A,Nelson E,Collins J E, et al.Characterization of Swine infertility and respiratory syndrome(SIRS) virus(isolate ATCC VR-2332)[J].J Vet Diagn Inves,1992,4(2):127-133.
[3] Cavanagh D.Nidovirales:a new order comprising Coronaviridae and Arteriviridae[J].Archives of Virology,1997,142(3):629-633.
[4] Den Boon J A,Faaberg K S,Meulenberg J J,et al.Processing and evolution of the N-terminal region of the arterivirus replicase ORF1a protein:identification of two papainlike cysteine proteases[J].Journal of Virology,1995,69(7):4500-4505.
[5] Snijder E J,Meulenberg J J.The molecular biology of arteriviruses[J].The Journal of General Virology,1998,79(Pt 5):961-979.
[6] Meulenberg J J,Petersen Den Besten A,De Kluyver E, et al.Molecular characterization of Lelystad virus[J].Veterinary Microbiology,1997,55(1/4):197-202.
[7] Beerens N,Selisko B,Ricagno S, et al.De novo initiation of RNA synthesis by the arterivirus RNA-dependent RNA polymerase[J].Journal of Virology,2007,81(16):8384-8395.
[8] Dong J,Zhang N,Ge X, et al.The interaction of nonstructural protein 9 with retinoblastoma protein benefits the replication of genotype 2Porcine reproductive and respiratory syndrome virus in vitro[J].Virology,2014,464-465:432-440.
[9] Li J,Guo D,Huang L,et al.The interaction between host Annexin A2 and viral Nsp9 is beneficial for replication of Porcine reproductive and respiratory syndrome virus[J].Virus Research,2014,189:106-113.
[10] Zhao S,Ge X,Wang X, et al.The DEAD-box RNA helicase 5 positively regulates the replication of Porcine reproductive and respiratory syndrome virus by interacting with viral Nsp9 in vitro[J].Virus Research,2015,195:217-224.
[11] Liu H,Wang Y,Duan H, et al.An intracellularly expressed Nsp9-specific nanobody in MARC-145 cells inhibits Porcine reproductive and respiratory syndrome virus replication[J].Veterinary Microbiology,2015,181(3/4):252-260.
[12] Gao L,Wang L,Huang C,et al.HP-PRRSV is attenuated by de-optimization of codon pair bias in its RNA-dependent RNA polymerase nsp9 gene[J].Virology,2015,485:135-144.
[13] Gao L,Zhang W,Sun Y,et al.Cryptoporus volvatus extract inhibits Porcine reproductive and respiratory syndrome virus(PRRSV) in vitro and in vivo[J].PLoS One,2013,8(5):e63767.
[14] Yang Q,Zhang Q,Tang J, et al.Lipid rafts both in cellular membrane and viral envelope are critical for PRRSV efficient infection[J].Virology,2015,484:170-180.
[15] Xie J,Zhou H,Cui J, et al.Inhibition of porcine reproductive and respiratory syndrome virus by specific siRNA targeting Nsp 9 gene[J].Infect Genet Evol,2014,28:64-70.
[16] 袁威,邓穗丹,邓又天,等.中药防制猪繁殖与呼吸综合征的研究进展[J].中国畜牧兽医,2013,40(2):193-197.
[17] Zhao M,Ning Z,Wang H,et al.Sequence analysis of NSP9 gene of 25 PRRSV strains from Guangdong province, subtropical southern China[J].Virus Genes,2013,46(1):88-96.
[18] 李嘉彬,赵孟孟,李玉谷,等.猪繁殖与呼吸综合征病毒变异株FS的分离鉴定与全基因组序列分析[J].中国畜牧兽医,2012,39(3):60-64.
[19] 赵孟孟,冯松林,王文佳, Nsp9 基因在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响[J].中国畜牧兽医,2016,43(12):3114-3120.
[20] Li Y,Zhou L,Zhang J, et al. Nsp9 and Nsp10 contribute to the fatal virulence of highly pathogenic Porcine reproductive and respiratory syndrome virus emerging in China[J].PLoS Pathogens,2014,10(7):e1004216.
[21] Burgara-Estrella A,Díaz I,Rodríguez-Gómez I M, et al.Predicted peptides from non-structural proteins of Porcine reproductive and respiratory syndrome virus are able to induce IFN-γ and IL-10[J].Viruses,2013,5(2):663-677.
[22] Parida R,Choi I S,Peterson D A,et al.Location of T-cell epitopes in nonstructural proteins 9 and 10 of type-Ⅱ Porcine reproductive and respiratory syndrome virus[J].Virus Research,2012,169(1):13-21.
[23] Tian D,Meng X J.Amino acid residues Ala283 and His421 in the RNA-dependent RNA polymerase of Porcine reproductive and respiratory syndrome virus play important roles in viral ribavirin sensitivity and quasispecies diversity[J].The Journal of General Virology,2016,97(1):53-59.
[24] An T Q,Zhou Y J,Liu G Q, et al.Genetic diversity and phylogenetic analysis of glycoprotein 5 of PRRSV isolates in mainland China from 1996 to 2006:coexistence of two NA-subgenotypes with great diversity[J].Veterinary Microbiology,2007,123(1/3):43-52.
[25] Zhou L,Chen S X,Zhang J L, et al.Molecular variation analysis of Porcine reproductive and respiratory syndrome virus in China[J].Virus Research,2009,145(1):97-105.
[26] Zhou L,Wang Z,Ding Y, et al.NADC30-like strain of Porcine reproductive and respiratory syndrome virus,China[J].Emerging Infectious Diseases,2015,21(12):2256-2257.

Please choose a citation manager

Content to export