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.
Institute of Molecular Biology and Biotechnology, Nik. Plastira 100, 71110 Heraklion, Crete, Greece. schinko@imbb.forth.gr
Johannes Benno
JB
Schinko
Kathrin
K
Hillebrand
Gregor
G
Bucher
D000818:Animals; D015870:Gene Expression; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D020218:Response Elements; D014231:Tribolium / Q000235:genetics* / Q000502:physiology
D006360:Heat-Shock Proteins; D019476:Insect Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins