ABSTRACT
Caenorhabditis elegans is a model organism that has been used to study human bacterial and viral pathogenesis. We report here the expression of human hepatitis delta viral antigens (HDAg) in C. elegans and measure the effect on the sterility, growth, and brood size in transgenic worms. Expression of HDAg under two different promoters, fib-1 (a ubiquitous promoter) and myo-2 (a pharynx-specific promoter), was achieved in C. elegans using dicistronic or tricistronic vectors derived from the operon CEOP5428. Transgenic worms expressing HDAg ubiquitously resulted in 20% to 70% sterility while those expressing HDAg in the pharynx displayed 70% sterility. Most of worms expressing HDAg in pharynx were arrested at larvae stage 2 or 3 and displayed a 70% reduction in brood size. Domain mapping experiments suggested that the nuclear localization signal of HDAg is required for the observed effect. Heat-shock induction of HDAg expression revealed that L4 larvae were the most sensitive to brood size reduction. These studies demonstrate that C. elegans can provide an additional model for studying HDAg interactions with host targets.
Subject(s)
Caenorhabditis elegans/growth & development , Caenorhabditis elegans/genetics , Hepatitis delta Antigens/metabolism , Reproduction/genetics , Animals , Clutch Size/genetics , DNA Primers/genetics , Fertility/genetics , Genetic Vectors/genetics , Humans , Microinjections , Microscopy , Promoter Regions, Genetic/genetics , RNA InterferenceABSTRACT
To meet the increasing need of simultaneously co-expressing two different genes in the same cell of transgenic Caenorhabditis elegans, here, we report the establishment of dicistronic vectors that contain an intercistronic region (ICR) of the C. elegans operon, CEOP5428. In these vectors, a green fluorescence protein (GFP) and a red FP (RFP) genes were placed in the first and second cistrons, respectively, which were separated by the ICR. Driven by the fibrillarin (fib-1) or myo-2 promoter, the GFP- and RFP-fusion proteins were consistently co-expressed in the entire worm cells or in the pharynx muscle cells of the transgenic worms, respectively. Our work demonstrates that ICR-containing dicistronic vectors could be developed into versatile co-expression systems in C. elegans for functional analysis of genes of interest.