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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

Analysis of PCR products of GX0101Δ(A+C)-BAC DNA. Lane M: DL2000 marker (TaKaRa Bio-Company, China); 1: the PCR product of GX0101Δ1 (A+C); 2: the PCR product of GX0101Δ (A+C). The bigger band demonstrated one of ORF (A+C) was replaced by kana gene (in lanes 1 and 2). The smaller band demonstrated the deletion of the second ORF(A+C) in GX0101Δ(A+C)-BAC DNA (in lane 2) compared to the smaller band that not deleted the second ORF(A+C) in GX0101Δ1(A+C)-BAC DNA (in lane 1).
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Figure 1: Analysis of PCR products of GX0101Δ(A+C)-BAC DNA. Lane M: DL2000 marker (TaKaRa Bio-Company, China); 1: the PCR product of GX0101Δ1 (A+C); 2: the PCR product of GX0101Δ (A+C). The bigger band demonstrated one of ORF (A+C) was replaced by kana gene (in lanes 1 and 2). The smaller band demonstrated the deletion of the second ORF(A+C) in GX0101Δ(A+C)-BAC DNA (in lane 2) compared to the smaller band that not deleted the second ORF(A+C) in GX0101Δ1(A+C)-BAC DNA (in lane 1).

Mentions: The deletion of the ORF(A+C) was confirmed by PCR with purified GX0101Δ1(A+C)-BAC and GX0101Δ(A+C)-BAC as templates [16]. As shown in Figure 1, the deletion of both copies of ORF(A+C) was confirmed by agarose gel electrophoresis of PCR. Then the GX0101Δ(A+C)-BAC DNA was transfected into CEF for the rescue of GX0101Δ(A+C) virus. As shown in Figure 2, the plaque size of GX0101Δ(A+C) was smaller than that of GX0101 at 96 h after infected in fresh CEF cells.


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)

Analysis of PCR products of GX0101Δ(A+C)-BAC DNA. Lane M: DL2000 marker (TaKaRa Bio-Company, China); 1: the PCR product of GX0101Δ1 (A+C); 2: the PCR product of GX0101Δ (A+C). The bigger band demonstrated one of ORF (A+C) was replaced by kana gene (in lanes 1 and 2). The smaller band demonstrated the deletion of the second ORF(A+C) in GX0101Δ(A+C)-BAC DNA (in lane 2) compared to the smaller band that not deleted the second ORF(A+C) in GX0101Δ1(A+C)-BAC DNA (in lane 1).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Analysis of PCR products of GX0101Δ(A+C)-BAC DNA. Lane M: DL2000 marker (TaKaRa Bio-Company, China); 1: the PCR product of GX0101Δ1 (A+C); 2: the PCR product of GX0101Δ (A+C). The bigger band demonstrated one of ORF (A+C) was replaced by kana gene (in lanes 1 and 2). The smaller band demonstrated the deletion of the second ORF(A+C) in GX0101Δ(A+C)-BAC DNA (in lane 2) compared to the smaller band that not deleted the second ORF(A+C) in GX0101Δ1(A+C)-BAC DNA (in lane 1).
Mentions: The deletion of the ORF(A+C) was confirmed by PCR with purified GX0101Δ1(A+C)-BAC and GX0101Δ(A+C)-BAC as templates [16]. As shown in Figure 1, the deletion of both copies of ORF(A+C) was confirmed by agarose gel electrophoresis of PCR. Then the GX0101Δ(A+C)-BAC DNA was transfected into CEF for the rescue of GX0101Δ(A+C) virus. As shown in Figure 2, the plaque size of GX0101Δ(A+C) was smaller than that of GX0101 at 96 h after infected in fresh CEF cells.

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