title: Construction and evaluation of a transgenic hsp16-GFP-lacZ Caenorhabditis elegans strain for environmental monitoring.
abstract: A novel integrated transgenic Caenorhabditis elegans strain (PC161) incorporates a double reporter construct with green fluorescent protein (GFP) and lacZ genes fused in-frame into the second exon of the hsp16-1 gene. This construct also includes the Simian Virus 40 (SV40) nuclear localization signal such that the fusion protein accumulates in the nuclei of expressing cells. The PC161 strain was used to monitor the effects of several known stressors, including heat, cadmium, and microwave radiation. The time course of induction was similar for both reporters but was strongly influenced by pretreatment conditions. The PC161 worms kept at 15 degrees C beforehand showed a steady increase in reporter expression (up to at least 16 h) when heated to 30 degrees C. However, if washed on ice prior to heat stress at 30 degrees C, PC161 worms showed a much steeper rise in reporter expression, reaching a maximum after 2.5 h and then plateauing. Heat shock induced strong expression of both reporter genes in all tissues apart from the germ line and early embryos. A highly significant linear dose-response relationship was observed for both transgenes with increasing cadmium concentrations (5-100 microg/ml). Prolonged exposure to microwave radiation (750 MHz and 0.5 W for 16 h) also induced expression of both transgenes at 25 and (to some extent) 27 degrees C, but only beta-galactosidase activity was detectable at 23 degrees C, and neither reporter was detectably expressed at 21 degrees C. Throughout all exposures, the lacZ reporter product was more readily detectable than coexpressed GFP. However, the GFP reporter affords opportunities to monitor the stress response in living worms.
authors.affiliation: Safety and Environmental Assurance Centre, Unilever Research Port Sunlight, Quarry Road East, Bebington, Wirral CH63 3JW, United Kingdom.
authors.forename: Helen E
authors.initials: HE
authors.lastname: David
authors.forename: Adam S
authors.initials: AS
authors.lastname: Dawe
authors.forename: David I
authors.initials: DI
authors.lastname: de Pomerai
authors.forename: Don
authors.initials: D
authors.lastname: Jones
authors.forename: E Peter M
authors.initials: EP
authors.lastname: Candido
authors.forename: Clare
authors.initials: C
authors.lastname: Daniells
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002104:Cadmium / Q000009:adverse effects; D017173:Caenorhabditis elegans / Q000235:genetics*; D004781:Environmental Exposure; D004784:Environmental Monitoring / Q000379:methods*; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006360:Heat-Shock Proteins / Q000235:genetics*; D007763:Lac Operon / Q000235:genetics*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D006360:Heat-Shock Proteins; D008164:Luminescent Proteins; D002104:Cadmium; D049452:Green Fluorescent Proteins