猪肾细胞源三种蛋白质对猪圆环病毒2型复制的调节作用

doi:10.11843/j.issn.0366-6964.2015.11.017

Abstract

Abstract:

To investigate the regulative effect of expression level of three proteins gene (AP2α2，Elf4 and ISCU) derived from porcine kidney cell line (PK-15) on porcine circovirus type 2 (PCV2) propagation efficiency,the genes of three proteins were amplified by RT-PCR and then three eukaryotic expression vectors were constructed.The RNAi fragments of these proteins were designed and cloned into pGPU6-GFP vector to construct shRNA vectors respectively.The efficiency of interference was determined by real-time PCR and three shRNA expression plasmids with higher efficiency of interference were determined.The PK-15 cells transfected with eukaryotic expression vectors or shRNA were screened by antibiotics of G418.The propagation efficiency of strains PCV2 LG in the cell lines was investigated using a capture enzyme-linked immunosorbent assay (ELISA) and an immunoperoxidase monolayer assay (IPMA).The results showed that Elf4 and ISCU overexpression increased PCV2 replication.In contrast，overexpression of AP2α2 exhibited no significant influence on PCV2 replication.In contrast，AP2α2，ISCU and Elf4 down-regulation by shRNA resulted in decreased PCV2 propagation，which demonstrated that these three proteins play a regulative role in PCV2 replication.

CLC Number:

S852.659.2

ZHANG Hui，TANG Qing-hai， HUANG Li-ping，WEI Yan-wu，LIU Chang-ming. Regulation Effect of Three Proteins Derived from Porcine Kidney Cells on Porcine Circovirus Type 2 Replication[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(11): 2040-2049.

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Regulation Effect of Three Proteins Derived from Porcine Kidney Cells on Porcine Circovirus Type 2 Replication

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