Core labeling of adenovirus with EGFP

Long P. Le, Helen N. Le, Amy R. Nelson, David A. Matthews, Masato Yamamoto, David T. Curiel

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

The study of adenovirus could greatly benefit from diverse methods of virus detection. Recently, it has been demonstrated that carboxy-terminal EGFP fusions of adenovirus core proteins Mu, V, and VII properly localize to the nucleus and display novel function in the cell. Based on these observations, we hypothesized that the core proteins may serve as targets for labeling the adenovirus core with fluorescent proteins. To this end, we constructed various chimeric expression vectors with fusion core genes (Mu-EGFP, V-EGFP, preVII-EGFP, and matVII-EGFP) while maintaining expression of the native proteins. Expression of the fusion core proteins was suboptimal using E1 expression vectors with both conventional CMV and modified (with adenovirus tripartite leader sequence) CMV5 promoters, resulting in non-labeled viral particles. However, robust expression equivalent to the native protein was observed when the fusion genes were placed in the deleted E3 region. The efficient Ad-wt-E3-V-EGFP and Ad-wt-E3-preVII-EGFP expression vectors were labeled allowing visualization of purified virus and tracking of the viral core during early infection. The vectors maintained their viral function, including viral DNA replication, viral DNA encapsidation, cytopathic effect, and thermostability. Core labeling offers a means to track the adenovirus core in vector targeting studies as well as basic adenovirus virology.

Original languageEnglish
Pages (from-to)291-302
Number of pages12
JournalVirology
Volume351
Issue number2
DOIs
StatePublished - Aug 1 2006
Externally publishedYes

Keywords

  • Adenovirus
  • Core proteins
  • EGFP
  • Fluorescent labeling
  • Gene therapy
  • Mu
  • V
  • VII
  • Vector

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    Le, L. P., Le, H. N., Nelson, A. R., Matthews, D. A., Yamamoto, M., & Curiel, D. T. (2006). Core labeling of adenovirus with EGFP. Virology, 351(2), 291-302. https://doi.org/10.1016/j.virol.2006.03.042