Identification of the core sequence elements in Penaeus stylirostris densovirus promoters

Arun K. Dhar, Krista N. Kaizer, Yelena M. Betz, Thomas N. Harvey, Dilip K. Lakshman

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

In silico analysis of three Penaeus stylirostris densovirus (PstDNV) promoters, designated P2, P11, and P61, revealed sequence motifs including the TATA box, downstream promoter element (DPE), GC- and A-rich regions, inverted repeat, activation sequence-1 like (ASL) box, and a conserved guanosine (G) at ?24. To delineate the regulatory role of these motifs on promoter activity, deletion constructs were made in a promoter assay vector, pGL3 Basic, that contains a luciferase reporter gene. Luciferase assay showed that P2 had the highest promoter activity followed by P11 and P61 in Sf9 cells. The deletions of inverted repeat, DPE, and GC-rich regions in P2 had the highest negative impact on this promoter. Deletions of DPE, G at the ?24, and ASL box in P11 had the highest negative impact on this promoter activity. In P61, DPE and G at ?24 are the two key regulators of transcriptional activity. Identification of the key transcriptional regulators is important in understanding the PstDNV pathogenesis in shrimp. This information is also valuable in constructing shrimp viral promoter-based vectors for protein expression in insect cell culture system as well as in shrimp.

Original languageEnglish (US)
Pages (from-to)367-375
Number of pages9
JournalVirus Genes
Volume43
Issue number3
DOIs
StatePublished - Dec 1 2011
Externally publishedYes

Keywords

  • IHHNV
  • Penaeus stylirostris densovirus
  • Promoter elements
  • Shrimp

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Virology

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