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An intron-containing,heat-inducible stress-70 gene in the millipede Tachypodoiulus niger (Julidae,Diplopoda)
Authors:Thomas Knigge  Lutz Bachmann  Heinz-R. Köhler
Affiliation:1. Laboratory of Ecotoxicology, EA 3222 PRES Normandie, Le Havre University, 25 Rue Philippe Lebon, F-76058, Le Havre Cedex, France
4. Laboratoire d’Ecotoxicologie, Université du Havre, 25, rue Ph. Lebon – CS 80540, F-76058, Le Havre Cedex, France
2. Department of Research and Collections, Natural History Museum, University of Oslo, P.O. Box 1172 Blindern, 0318, Oslo, Norway
3. Animal Physiological Ecology, Institute of Evolution and Ecology, University of Tübingen, Konrad-Adenauer-Str. 20, D-72072, Tübingen, Germany
Abstract:The highly conserved part of the nucleotide-binding domain of the hsp70 gene family was amplified from the soil diplopod Tachypodoiulus niger (Julidae, Diplopoda). Genomic DNA yielded 701, 549 and 540 bp sequences, whereas cDNA from heat shocked animals produced only one distinct fragment of 543 bp. The sequences could be classified as a 70 kDa heat shock protein (hsp70), the corresponding 70 kDa heat shock cognate (hsc70) and a glucose-related hsp70 homologue (grp78). Comparisons of genomic and cDNA sequences of hsc70 identified two introns within the consensus sequence. Generally, stress-70 expression levels were low, which hampered successful RT-PCR and subsequent subcloning. Following experimental heat shock, however, the spliced hsc70 was amplified predominantly, instead of its inducible homologue hsp70. This finding suggests that microevolution in this soil-dwelling arthropod is directed towards low constitutive stress-70 levels and that the capacity for stress-70 induction presumably is limited. hsc70, albeit having introns, apparently is inducible and contributes to the stress-70 response.
Keywords:Stress response   Soil invertebrate   Arthropods   Heat shock proteins   Heat shock cognates   Glucose-related proteins   Biomarker
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