Porcine Interferon Stimulated Gene 12a Restricts Porcine Reproductive and Respiratory Syndrome Virus Replication in MARC-145 Cells
Abstract
:1. Introduction
2. Results
2.1. Porcine ISG12A Sequence and Phylogenetic Analysis
2.2. Porcine ISG12A Is Located in Mitochondria and Regulate Cell Cycle
2.3. Increased Expression of Porcine ISG12A after PRRSV Challenge In Vivo
2.4. Porcine ISG12A Restricted PRRSV Replication in MARC-145 Cells
2.5. Knockdown of Endogenous ISG12A Enhanced PRRSV Replication in MARC-145 Cells
3. Discussion
4. Materials and Methods
4.1. Cells, Tissues, and Virus
4.2. Sequence Analysis
4.3. Expression Vectors
4.4. Confocal Microscopy
4.5. Flow Cytometry
4.6. Gene Expression in Cells and Tissues during PRRSV Challenge In Vivo
4.7. Transient Transfection and Real-Time Quantitative PCR
4.8. Immunofluorescence Assay
4.9. Statistical Tests
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Gene | Primer Sequence (5′–3′) | T (°C) | Application |
---|---|---|---|
pISG12A-PF1 | ATCTCGAGTATGGCCATGGTAGCCACCC | 58 | Expression vector construction |
pISG12A-PR1 | GTAAGCTTCAAGCCCACTATTTTGGTAC | ||
pISG12A-PF2 | GCCACCCAAGCAGTCTTATCCTC | 60 | qRT-PCR |
pISG12A-PR2 | TGTCCCCAGCAAGGATCCCA | ||
mISG12A-PF3 | GTGGCCTCCGCTTTCACCT | 60 | qRT-PCR |
mISG12A-PR3 | TTCCCGTCGCAGTGAAACCC | ||
ORF7-PF | CCCATTTCCCTCTAGCGACT | 60 | qRT-PCR |
ORF7-PR | GCGTCGGCAAACTAAACTCCA | ||
mGAPDH-PF | TGGGGAAGGTGAAGGTCGG | 60 | qRT-PCR |
mGAPDH-PR | TCCTGGAAGATGGTGATGGG | ||
pGAPDH-PF | CGTCCCTGAGACACGATGGT | 60 | qRT-PCR |
pGAPDH-PR | GCCTTGACTGTGCCGTGGAAT | ||
ISG12A-siRNA | GCUUUCACCUCAUCAGCAGUG | Macaque ISG12A siRNA | |
NC-siRNA | UUCUUCGAACGUGUCACGUTT | Negative control siRNA |
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Ji, L.; Zhou, X.; Liang, W.; Liu, J.; Liu, B. Porcine Interferon Stimulated Gene 12a Restricts Porcine Reproductive and Respiratory Syndrome Virus Replication in MARC-145 Cells. Int. J. Mol. Sci. 2017, 18, 1613. https://doi.org/10.3390/ijms18081613
Ji L, Zhou X, Liang W, Liu J, Liu B. Porcine Interferon Stimulated Gene 12a Restricts Porcine Reproductive and Respiratory Syndrome Virus Replication in MARC-145 Cells. International Journal of Molecular Sciences. 2017; 18(8):1613. https://doi.org/10.3390/ijms18081613
Chicago/Turabian StyleJi, Likai, Xiang Zhou, Wan Liang, Jianjian Liu, and Bang Liu. 2017. "Porcine Interferon Stimulated Gene 12a Restricts Porcine Reproductive and Respiratory Syndrome Virus Replication in MARC-145 Cells" International Journal of Molecular Sciences 18, no. 8: 1613. https://doi.org/10.3390/ijms18081613