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Identification of Virus Encoding MicroRNAs Using 454 FLX Sequencing Platform

  • Byung-Whi KongEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 733)

Abstract

MicroRNAs are a class of small noncoding RNA molecules that play a pivotal role in the regulation of gene expression at the posttranscriptional level. Most large double-stranded DNA viruses, mainly the herpesvirus family, are known to express miRNAs. Viral miRNAs can regulate both viral- and cellular transcripts. By eliminating cloning steps for large number of Sanger sequencing reactions, recent development of massively parallel next-generation sequencing methods has accelerated identification of small RNA species expressed from viruses, prokaryotes, and eukaryotes. The miRNAs expressed from infectious laryngotracheitis virus (ILTV), which is an alphaherpesvirus belonging to the herpesviridae family and which causes an acute respiratory disorder in chicken, were identified by small RNA enrichment and the 454 FLX sequencing method.

Key words

Viral microRNA Herpesvirus FLX sequencing 

Notes

Acknowledgments

We thank Dr. Joon Jin Song, University of Arkansas, for the support in sorting sequence reads. This work was supported by Arkansas Agricultural Experimental Station.

References

  1. 1.
    Bushati, N.and Cohen, S.M. (2007) microRNA functions. Annu Rev Cell Dev Biol 23, 175  –205PubMedCrossRefGoogle Scholar
  2. 2.
    Cullen, B.R. (2006) Viruses and microRNAs. Nat Genet 38 Suppl, S25–30Google Scholar
  3. 3.
    Burnside, J., Ouyang, M., Anderson, A., Bernberg, E., Lu, C., Meyers, B.C., Green, P.J., Markis, M., Isaacs, G., Huang, E., and Morgan, R.W. (2008) Deep sequencing of chicken microRNAs. BMC Genomics 9, 185PubMedCrossRefGoogle Scholar
  4. 4.
    Waidner, L.A., Morgan, R.W., Anderson, A.S., Bernberg, E.L., Kamboj, S., Garcia, M., Riblet, S.M., Ouyang, M., Isaacs, G.K., Markis, M., Meyers, B.C., Green, P.J., and Burnside, J. (2009) MicroRNAs of Gallid and Meleagrid herpesviruses show generally ­conserved genomic locations and are ­virus-specific. Virology 388, 128  –36PubMedCrossRefGoogle Scholar
  5. 5.
    Rachamadugu, R., Lee, J.Y., Wooming, A., and Kong, B.W. (2009) Identification and expression analysis of infectious laryngotracheitis virus encoding microRNAs. Virus Genes Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Poultry Science, and Cell and Molecular Biology ProgramUniversity of ArkansasFayettevilleUSA

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