Differentiation of Oncogenic and Nononcogenic Strains of Marek's Disease Virus Type 1 by Using Polymerase Chain Reaction DNA Amplification

Zhu, Geng-Sheng; Ojima, Tomoko; Hatori, Takashi; Ihara, Takeshi; Mizukoshi, Noriko; Kato, Atsushi; Ueda, Seinosuke; Hirai, Kanji

doi:10.2307/1591759

Cited by 40 publications

(18 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…MD commonly appears in 3-4 week old chickens and gradually builds to a peak between 12 and 30 weeks of age. Polymerase chain reaction (PCR) has emerged in recent years as an additional diagnostic tool offering the advantages of serotype specificity and the ability to differentiate between vaccinal and field strains of MDV serotype-1 [4,8,12]. The importance of BamHI-H region in the pathogenesis of MDV was initially pointed out; when it was shown that there was an increase in the number of copies of 132 bp repeats within the BamHI-H region of inverted repeats (IR) as a result of attenuation by cell culture passage [9].…”

mentioning

confidence: 99%

See 1 more Smart Citation

Characterization of 132 bp Repeats BamH1-H Region in Pathogenic Marek’s Disease Virus of Poultry in Gujarat, India, Using PCR and Sequencing

et al. 2011

View full text Add to dashboard Cite

A total of 34 clinical samples and four Marek's disease virus (MDV) vaccines were tested using primer BamH1/BamH2 in layer birds of poultry. Out of 34 samples tested for detection of MDV, 32 samples produced approximately 434 bp product. All the three HVT vaccines as well as SB-1 (MDV-2) vaccine failed to produce the expected amplicons, there by proving negative for the targeted 132 bp repeats of MDV genome by the primers BamH1/BamH2. Resultant PCR products of the field samples were purified and sequenced and resulted in 378 bp long sequences. PCR was found very satisfactory in detecting the presence of MDV either in feather follicle or in tissue samples. Sequencing study has proved beyond doubt that the two representative samples contained two 132 bp repeats indicating the virulent nature of the field virus.

show abstract

mentioning

confidence: 99%

“…amplification of 132 bp repeats) in MDV type-1 as an indicator for critical genomic rearrangement leading to the attenuation of virus virulence. In a similar study, Zhu et al [12] differentiated the oncogenic and non-oncogenic strain of MDV-1 by primer chosen from the sequence within the long inverted repeats of MDV-1 DNA.…”

mentioning

confidence: 99%

Characterization of 132 bp Repeats BamH1-H Region in Pathogenic Marek’s Disease Virus of Poultry in Gujarat, India, Using PCR and Sequencing

et al. 2011

View full text Add to dashboard Cite

show abstract

“…This transcript is mapped in the internal repeat long (IRL) region of the MDV genome containing the tandem repeats of the 132-bp repeat region. This repeat region is expanded during attenuation by in vitro passage and also found in vaccine strain CVI988 [21,22]. Recently, it was shown that the expansion of the 132-bp repeat is not sufficient in itself to attenuate pathogenic MDV [23].…”

Section: Introductionmentioning

confidence: 99%

Marek’s disease virus unique genes pp38 and pp24 are essential for transactivating the bi-directional promoters for the 1.8 kb mRNA transcripts

2007

View full text Add to dashboard Cite

The pp38 and pp24 genes of Marek's diseases virus (MDV) share the same promoter, which controls the transcription of pp38 or pp24 and a 1.8-kb mRNA bi-directionally. To understand the trans-activating activity of pp38 and pp24 on the bi-directional promoter, both genes were cloned into pcDNA-3 or pBudCE4.1 vectors either singly or in combination. These plasmids were expressed in transfected chicken embryonic fibroblast (CEF) cells. Chloramphenicol acetyltransferase (CAT) activity expressed under the control of the promoter in CEF co-transfected with pP(1.8 kb)-CAT and pBud-pp38-pp24 was significantly higher than that following transfection with only pBud-pp38 or pBud-pp24. This indicates the combination of pp24 and pp38 together are essential for the activation of the promoter. In DNA mobility shift assays, the promoter binds to pp38 and pp24 together, but not to pp38 or pp24 alone. By competitive inhibition tests with a set of DNA fragments from the promoter region, the sequence 5'-CTGCTCATTT-3' was identified as the core sequence for binding by pp38-pp24 in up-regulation of the bi-directional promoter activity.

show abstract

“…This repeat region is expanded during in vitro passage and the 1.8-kb mRNA becomes disrupted and heterogeneous [9]. In virulent oncogenic strains of MDV, there are only two copies of the 132-bp but are multiple copies up to 100 in attenuated viruses [16,17]. For a very long time, it was suggested that the expansion of the 132-bp region, disrupts the 1.8-kb mRNA that is essential for oncogenicity, thereby resulting in attenuation [9,18].…”

Section: Introductionmentioning

confidence: 99%

The Role of pp38 in Regulation of Marek’s Disease Virus Bi-directional Promoter between pp38 and 1.8-kb mRNA

Ding

Cui

Lee³

et al. 2006

Virus Genes

View full text Add to dashboard Cite

Marek's disease virus (MDV) contains a bi-directional promoters located between pp38 gene and 1.8-kb mRNA in the long inverted repeat region of the viral genome. The involvement of pp38 gene in up-regulating the activity of these promoters was analyzed by transient expression of chloramphenicol acetyltransferase (CAT) reporter gene. Two CAT reporter plasmids, pP(pp38)-CAT and pP(1.8-kb)-CAT, were constructed to express CAT under the control of the bi-directional promoter in both orientations. These plasmids were transfected into chicken embryonic fibroblast (CEF), infected with rMd5 and pp38 deleted rMd5 (rMd5/Deltapp38), respectively. No CAT activity was detected in uninfected CEF as expected. CAT activities in rMd5/Deltapp38 virus infected CEF (rMd5/Deltapp38-CEF) were 3.5-fold lower using pP(pp38)-CAT and 12-fold lower using pP(1.8-kb)-CAT than those of the parental rMd5 infected CEF (rMd5-CEF). The significantly lower promoter activity in the pp38 deletion virus suggests that pp38 can regulate the activity of the bi-directional promoters, especially in the direction of 1.8-kb mRNA family. Co-transfection of pp38-expressing plasmid (pcDNA-pp38) into rMd5/Deltapp38-CEF significantly increased the activity of the bi-directional promoters using either pP(pp38)-CAT or pP(1.8-kb)-CAT. DNA mobility shift assay showed a binding of the 73-bp sequence of the bi-directional promoter with rMd5-CEF but not with rMd5/Deltapp38-CEF or uninfected CEF lysates. However, rMd5/Deltapp38-CEF lysates could bind the same 73-bp promoter sequence when co-transfected with pp38-expressing plasmid (pcDNA-pp38). All these data taken together suggest pp38 plays an important role in regulating the transcriptional activity of the bi-directional promoter.

show abstract

Differentiation of Oncogenic and Nononcogenic Strains of Marek's Disease Virus Type 1 by Using Polymerase Chain Reaction DNA Amplification

Cited by 40 publications

References 21 publications

Characterization of 132 bp Repeats BamH1-H Region in Pathogenic Marek’s Disease Virus of Poultry in Gujarat, India, Using PCR and Sequencing

Characterization of 132 bp Repeats BamH1-H Region in Pathogenic Marek’s Disease Virus of Poultry in Gujarat, India, Using PCR and Sequencing

Marek’s disease virus unique genes pp38 and pp24 are essential for transactivating the bi-directional promoters for the 1.8 kb mRNA transcripts

The Role of pp38 in Regulation of Marek’s Disease Virus Bi-directional Promoter between pp38 and 1.8-kb mRNA

Contact Info

Product

Resources

About