Abstract
Immediate-early (IE) genes are the first class of viral genes expressed after primary infection or reactivation. As transcription of IE genes does not require prior viral protein synthesis, this class of genes is experimentally defined by their transcription following primary infection or reactivation in the presence of inhibitors of protein synthesis. This chapter describes an approach to identify IE genes in a novel herpesvirus genome. Transcription of IE genes is selectively induced with sodium butyrate in the presence of the protein synthesis inhibitor cycloheximide. The transcripts of the induced genes are identified by using a cDNA subtraction-based method of gene expression screening.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Roizman, B. and Pellett, P. (2002) The family Herpesviridae: a brief introduction, in Fields Virology, 4th ed. (Knipe, D. M., Howley, P. M., Griffin, D. E., et al., eds.), Lippincott Williams & Wilkins, Philadelphia, pp. 2381–2397.
Miller, G. (1990) The switch between latency and replication of Epstein-Barr virus. J. Infect. Dis. 161, 833–844.
Wu, F. Y., Chen, H., Wang, S. E., et al. (2003) CCAAT/enhancer binding protein alpha interacts with ZTA and mediates ZTA-induced p21CIP-1 accumulation and G1 cell cycle arrest during the Epstein-Barr virus lytic cycle. J. Virol. 77, 1481–1500.
Morrison, T. E., Mauser, A., Wong, A., Ting, J. P., and Kenney, S. C. (2001) Inhibition of IFN-gamma signaling by an Epstein-Barr virus immediate-early protein. Immunity 15, 787–799.
Honess, R. W. and Roizman, B. (1974) Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. J. Virol. 14, 8–19.
Wathen, M. W., Thomsen, D. R., and Stinski, M. F. (1981) Temporal regulation of human cytomegalovirus transcription at immediate early and early times after infection. J. Virol. 38, 446–459.
Zhu, F. X., Cusano, T., and Yuan, Y. (1999) Identification of the immediate-early transcripts of Kaposi’s sarcoma-associated herpesvirus. J. Virol. 73, 5556–5567.
Ebrahimi, B., Dutia, B. M., Roberts, K. L., et al. (2003) Transcriptome profile of murine gammaherpesvirus-68 lytic infection. J. Gen. Virol. 84, 99–109.
Cesarman, E., Moore, P. S., Rao, P. H., Inghirami, G., Knowles, D. M., and Chang, Y. (1995) In vitro establishment and characterization of two acquired immunodeficiency syndrome-related lymphoma cell lines (BC-1 and BC-2) containing Kaposi’s sarcoma-associated herpesvirus-like (KSHV) DNA sequences. Blood 86, 2708–2714.
Renne, R., Zhong, W., Herndier, B., et al. (1996) Lytic growth of Kaposi’s sarcoma-associated herpesvirus (human herpesvirus 8) in culture. Nat. Med. 2,342–346.
Wang, Z. and Brown, D. D. (1991) A gene expression screen. Proc. Natl.Acad. Sci. USA 88, 11505–11509.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Yuan, Y. (2005). Identification and Characterization of Herpesviral Immediate-Early Genes. In: Lieberman, P.M. (eds) DNA Viruses. Methods in Molecular Biology, vol 292. Humana Press. https://doi.org/10.1385/1-59259-848-X:231
Download citation
DOI: https://doi.org/10.1385/1-59259-848-X:231
Publisher Name: Humana Press
Print ISBN: 978-1-58829-353-4
Online ISBN: 978-1-59259-848-9
eBook Packages: Springer Protocols