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Deletion of 1.8-kb mRNA of Marek's disease virus decreases its replication ability but not oncogenicity.

Sun A, Li Y, Wang J, Su S, Chen H, Zhu H, Ding J, Cui Z - Virol. J. (2010)

Bottom Line: Based on the BAC clone of GX0101, the 1.8-kb mRNA deletion mutant GX0101Δ(A+C) was constructed.Further studies in vitro demonstrated that deletion of 1.8-kb mRNA significantly decreased the transcriptional activity of the bi-directional promoter between 1.8-kb mRNA and pp38 genes of MDV.These results suggested that the 1.8-kb mRNA did not directly influence the oncogenesis but related to the replication ability of MDV.

View Article: PubMed Central - HTML - PubMed

Affiliation: Animal Science and Technology College, Shandong Agricultural University, Tai'an, Shandong 271018, China.

ABSTRACT

Background: The 1.8-kb mRNA was reported as one of the oncogenesis-related genes of Marek's disease virus (MDV). In this study, the bacterial artificial chromosome (BAC) clone of a MDV field strain GX0101 was used as the platform to generate mutant MDV to examine the functional roles of 1.8-kb mRNA.

Results: Based on the BAC clone of GX0101, the 1.8-kb mRNA deletion mutant GX0101Δ(A+C) was constructed. The present experiments indicated that GX0101Δ(A+C) retained a low level of oncogenicity, and it showed a decreased replication capacity in vitro and in vivo when compared with its parent virus, GX0101. Further studies in vitro demonstrated that deletion of 1.8-kb mRNA significantly decreased the transcriptional activity of the bi-directional promoter between 1.8-kb mRNA and pp38 genes of MDV.

Conclusion: These results suggested that the 1.8-kb mRNA did not directly influence the oncogenesis but related to the replication ability of MDV.

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Related in: MedlinePlus

Growth curves of GX0101Δ(A+C) and GX0101 in vitro. 100 PFU GX0101 and GX0101Δ(A+C) were inoculated onto six-well plates seeded with 2×106 CEFs and incubated at 37°C, 5% CO2, respectively. At hours 0, 24, 48, 72, 96, 120 and 144 p.i., the infected cells were trypsinized and serial 10-fold dilutions were added onto six-well plates of CEFs, visible viral plaques were counted on days 5 p.i. by IFA. The means ± SD at each time point were shown, *P < 0.05 compared with those in GX0101 group. It was demonstrated that the mutant virus GX0101Δ(A+C) exhibited a decreased replication ability in CEF compared with GX0101 at hours 72, 96, 120 and 144 p.i. (P < 0.05).
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Figure 3: Growth curves of GX0101Δ(A+C) and GX0101 in vitro. 100 PFU GX0101 and GX0101Δ(A+C) were inoculated onto six-well plates seeded with 2×106 CEFs and incubated at 37°C, 5% CO2, respectively. At hours 0, 24, 48, 72, 96, 120 and 144 p.i., the infected cells were trypsinized and serial 10-fold dilutions were added onto six-well plates of CEFs, visible viral plaques were counted on days 5 p.i. by IFA. The means ± SD at each time point were shown, *P < 0.05 compared with those in GX0101 group. It was demonstrated that the mutant virus GX0101Δ(A+C) exhibited a decreased replication ability in CEF compared with GX0101 at hours 72, 96, 120 and 144 p.i. (P < 0.05).

Mentions: To determine whether the deletion of the 1.8-kb mRNA had any effect on GX0101Δ(A+C) growth replication in vitro, the growth rate of GX0101Δ(A+C) virus was compared with that of GX0101. As shown in Figure 3, it was demonstrated that the recombinant virus GX0101Δ(A+C) exhibited a decreased replication ability in CEF compared with GX0101 at hours 72, 96, 120 and 144 post inoculation (p.i.) (P < 0.05).


Deletion of 1.8-kb mRNA of Marek's disease virus decreases its replication ability but not oncogenicity.

Sun A, Li Y, Wang J, Su S, Chen H, Zhu H, Ding J, Cui Z - Virol. J. (2010)

Growth curves of GX0101Δ(A+C) and GX0101 in vitro. 100 PFU GX0101 and GX0101Δ(A+C) were inoculated onto six-well plates seeded with 2×106 CEFs and incubated at 37°C, 5% CO2, respectively. At hours 0, 24, 48, 72, 96, 120 and 144 p.i., the infected cells were trypsinized and serial 10-fold dilutions were added onto six-well plates of CEFs, visible viral plaques were counted on days 5 p.i. by IFA. The means ± SD at each time point were shown, *P < 0.05 compared with those in GX0101 group. It was demonstrated that the mutant virus GX0101Δ(A+C) exhibited a decreased replication ability in CEF compared with GX0101 at hours 72, 96, 120 and 144 p.i. (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2984594&req=5

Figure 3: Growth curves of GX0101Δ(A+C) and GX0101 in vitro. 100 PFU GX0101 and GX0101Δ(A+C) were inoculated onto six-well plates seeded with 2×106 CEFs and incubated at 37°C, 5% CO2, respectively. At hours 0, 24, 48, 72, 96, 120 and 144 p.i., the infected cells were trypsinized and serial 10-fold dilutions were added onto six-well plates of CEFs, visible viral plaques were counted on days 5 p.i. by IFA. The means ± SD at each time point were shown, *P < 0.05 compared with those in GX0101 group. It was demonstrated that the mutant virus GX0101Δ(A+C) exhibited a decreased replication ability in CEF compared with GX0101 at hours 72, 96, 120 and 144 p.i. (P < 0.05).
Mentions: To determine whether the deletion of the 1.8-kb mRNA had any effect on GX0101Δ(A+C) growth replication in vitro, the growth rate of GX0101Δ(A+C) virus was compared with that of GX0101. As shown in Figure 3, it was demonstrated that the recombinant virus GX0101Δ(A+C) exhibited a decreased replication ability in CEF compared with GX0101 at hours 72, 96, 120 and 144 post inoculation (p.i.) (P < 0.05).

Bottom Line: Based on the BAC clone of GX0101, the 1.8-kb mRNA deletion mutant GX0101Δ(A+C) was constructed.Further studies in vitro demonstrated that deletion of 1.8-kb mRNA significantly decreased the transcriptional activity of the bi-directional promoter between 1.8-kb mRNA and pp38 genes of MDV.These results suggested that the 1.8-kb mRNA did not directly influence the oncogenesis but related to the replication ability of MDV.

View Article: PubMed Central - HTML - PubMed

Affiliation: Animal Science and Technology College, Shandong Agricultural University, Tai'an, Shandong 271018, China.

ABSTRACT

Background: The 1.8-kb mRNA was reported as one of the oncogenesis-related genes of Marek's disease virus (MDV). In this study, the bacterial artificial chromosome (BAC) clone of a MDV field strain GX0101 was used as the platform to generate mutant MDV to examine the functional roles of 1.8-kb mRNA.

Results: Based on the BAC clone of GX0101, the 1.8-kb mRNA deletion mutant GX0101Δ(A+C) was constructed. The present experiments indicated that GX0101Δ(A+C) retained a low level of oncogenicity, and it showed a decreased replication capacity in vitro and in vivo when compared with its parent virus, GX0101. Further studies in vitro demonstrated that deletion of 1.8-kb mRNA significantly decreased the transcriptional activity of the bi-directional promoter between 1.8-kb mRNA and pp38 genes of MDV.

Conclusion: These results suggested that the 1.8-kb mRNA did not directly influence the oncogenesis but related to the replication ability of MDV.

Show MeSH
Related in: MedlinePlus