Generation of PCV2 in PK15 cells transfected with recombinant baculovirus containing a 1.1 copy of the PCV2 genome

Cai, Jie; Xie, Xiaohong; Hu, Yi; Zhan, Yang; Yu, Wanting; Wang, Aibing; Wang, Naidong

doi:10.1556/004.2017.027

Cited by 3 publications

(2 citation statements)

References 36 publications

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“…PK-15 cells free of PCV2 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), and they were cultured in Dulbecco’s modified Eagle medium (HyClone, GE, Beijing, China) supplemented with 10% fetal bovine serum (HyClone, GE, Beijing, China). The cell line was used because it is a widely accepted in vitro infection model, especially in the field of PCV2 virology, from which underlying molecular and cellular mechanisms of PCV2 infection have been learned [ 26 , 27 , 28 , 29 , 30 , 31 ]. The PCV2 strain 17HNA1 (Accession no.…”

Section: Methodsmentioning

confidence: 99%

Identification of lncRNAs Involved in PCV2 Infection of PK-15 Cells

Leng

Pan

et al. 2020

Pathogens

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Porcine circovirus type 2 (PCV2) can cause severe disease in infected pigs, resulting in massive economic loss for the swine industry. Transcriptomic and proteomic approaches have been widely employed to identify the underlying molecular mechanisms of the PCV2 infection. Numerous differentially expressed mRNAs, miRNAs, and proteins, together with their associated signaling pathways, have been identified during PCV2 infection, paving the way for analysis of their biological functions. Long noncoding RNAs (lncRNAs) are important regulators of multiple biological processes. However, little is known regarding their role in the PCV2 infection. Hence, in our study, RNA-seq was performed by infecting PK-15 cells with PCV2. Analysis of the differentially expressed genes (DEGs) suggested that the cytoskeleton, apoptosis, cell division, and protein phosphorylation were significantly disturbed. Then, using stringent parameters, six lncRNAs were identified. Additionally, potential targets of the lncRNAs were predicted using both cis- and trans-prediction methods. Interestingly, we found that the HOXB (Homeobox B) gene cluster was probably the target of the lncRNA LOC106505099. Enrichment analysis of the target genes showed that numerous developmental processes were altered during PCV2 infection. Therefore, our study revealed that lncRNAs might affect porcine embryonic development through the regulation of the HOXB genes.

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Section: Methodsmentioning

confidence: 99%

Identification of lncRNAs Involved in PCV2 Infection of PK-15 Cells

Leng

Pan

et al. 2020

Pathogens

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“…To construct the infectious clones, a set of primers including HY008, HY009, HY010, HY011 for pSP72-PCV2b-1B-YiY-3-2-1 (IF-YiY-3-2-1) and pSP72-PCV2b-1B-YiY-3-2-3 (IF-YiY-3-2-3), a set of primers containing HY010, HY011, HY012, HY013 for pSP72-PCV2d-YiY-3-2-10 (IF-YiY-3-2-10), were designed and synthesized to amplify the DNA fragment of 1.1 copy PCV2 genome via PCR (Table 1 and Fig. 1) [27]. PCR products were separated on 1% agarose gels by electrophoresis.…”

Section: Methodsmentioning

confidence: 99%

Truncated Rep protein of porcine circovirus 2 (PCV2) caused by a naturally occurring mutation reduced virus replication in PK15 cells

Cai

Zhan³

et al. 2019

BMC Vet Res

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Background Porcine circovirus 2 (PCV2) is the causative agent of porcine circovirus-associated diseases (PCVADs). The infection of PCV2 is widespread and has serious consequence, thereby causing significant economic losses in the swine industry worldwide. Previously, we found that a strain named YiY-3-2-3 has a naturally occurring point mutation (G 710 to A 710 ) in ORF1 region, which leads to a shorten product of the rep gene (945 to 660 base pair). Importantly, the Rep protein is responsible for genome replication of PCV2. To explore the effects of this mutation on the PCV2 replication, in the current study we constructed infectious clone of this IF-YiY-3-2-3, as well as those of its two parental strains of IF-YiY-3-2-1 and IF-YiY-3-2-10. Subsequently, these infectious clones which have 1.1 copy of PCV2 genome of their corresponding strains were transfected into PK15 cells to obtain rescued viruses, respectively. Results Though all of the three infectious clones could be rescued, the copy number and infectivity of these rescued viruses were significantly different, as analyzed by fluorescence quantitative PCR, Tissue culture infectious dose 50 (TCID 50 ), and indirect immunofluorescence assay (IFA). Notably, whether the PCV2 copy number, viral titer or the infectivity of rescued viruses from infectious clone IF-YiY-3-2-3 was significantly less than those of its parental clones. Meanwhile, the spatial structure of the Rep protein from the IF-YiY-3-2-3 displayed an apparent truncation at the C-terminal. Conclusions These findings therefore suggest that the Rep protein with truncated C-terminal would reduce virus replication and infectivity, and there might also exist both favorable and unfavorable mutations in the ORF1 of PCV2 in the process of its evolution.

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