Heat shock-mediated misexpression of genes in the beetle Tribolium castaneum
Insect gene function has mainly been studied in the fruit fly Drosophila melanogaster because in this species many techniques and resources are available for gene knock down and the ectopic activation of gene function. However, in order to study biological aspects that are not represented by the Drosophila model, and in order to test to what degree gene functions are conserved within insects and what changes in gene function accompanied the evolution of novel traits, the establishment of respective tools in other insect species is required. While gene knock down can be induced by RNA interference in many insects, methods to misexpress genes are much less developed. In order to allow misexpression of genes in a timely controlled manner in the red flour beetle Tribolium castaneum, we have established a heat shock-mediated misexpression system. We show that endogenous heat shock elements perform better than artificial heat shock elements derived from vertebrates. We carefully determine the optimal conditions for heat shock and define a core promoter for use in future constructs. Finally, using this system, we study the effects of misexpressing the head patterning gene Tc-orthodenticle1 (Tc-otd1), We show that Tc-otd1 suppresses Tc-wingless (Tc-wg) in the trunk and to some degree in the head.
KeywordsHeat shock hsp68 hsp70 Misexpression Orthodenticle
We thank Dr. Thomas Czerny for the artificial heat shock elements, Dr. Fakrudin Bashasab for identifying the hsp68 promoter. Dr. Michael Schoppmeier and Dr. Jochen Trauner for help with the interpretation of the signal in the ovaries. We also thank Anna Gilles for help with in situ hybridizations and helpful discussions and Dr. Andrew D. Peel for language corrections.
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