Functional analysis of putative operons in Brugia malayi

Canhui Liu, Ana Oliveira, Chitra Chauhan, Elodie Ghedin, Thomas R. Unnasch

Research output: Contribution to journalArticle

Abstract

Operons are a common mode of gene organization in Caenorhabditis elegans. Similar gene arrangements suggest that functional operons may exist in Brugia malayi. To definitively test this hypothesis, a bicistronic reporter vector consisting of an upstream firefly luciferase gene and a downstream renilla luciferase gene was constructed. The genome was then surveyed to identify 15 gene pairs that were likely to represent operons. Two of four domains upstream of the 5′ gene from these clusters exhibited promoter activity. When constructs replicating the promoter and intergenic arrangement found in the native putative operon were transfected into embryos, both firefly and renilla activities were detected, while constructs with the promoter alone or intergenic region alone produced no activity from the downstream reporter. These data confirm that functional operons exist in B. malayi. Mutation of three U-rich element homologues present in one of the operons resulted in a decrease in downstream renilla reporter activity, suggesting that these were important in mRNA maturation. Hemi-nested reverse transcriptase-PCR assays demonstrated that while the mRNA encoding the native downstream open reading frame of one operon contained an SL1 spliced leader at its 5′ end, the renilla gene mRNA produced from the corresponding transgenic construct did not.

Original languageEnglish (US)
Pages (from-to)63-71
Number of pages9
JournalInternational Journal for Parasitology
Volume40
Issue number1
DOIs
StatePublished - Jan 2010

Keywords

  • Biolositics
  • Filariasis
  • Operon
  • Trans-splicing
  • Transfection

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

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