Institute of Molecular Biology and Biotechnology, Nik. Plastira 100, 71110 Heraklion, Crete, Greece.
Dev Genes Evol. 2012 Sep;222(5):287-98. doi: 10.1007/s00427-012-0412-x. Epub 2012 Jul 20.
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.
昆虫基因功能主要在黑腹果蝇 Drosophila melanogaster 中进行研究，因为在这个物种中，有许多技术和资源可用于基因敲除和异位激活基因功能。然而，为了研究果蝇模型未涵盖的生物学方面，以及为了测试基因功能在昆虫内部的保守程度以及基因功能的哪些变化伴随着新特征的进化，需要在其他昆虫物种中建立各自的工具。虽然许多昆虫中可以通过 RNA 干扰诱导基因敲除，但基因过表达的方法则发展得较少。为了允许在红粉甲虫 Tribolium castaneum 中及时进行基因的时间控制过表达，我们建立了一种热休克介导的过表达系统。我们发现内源性热休克元件比来自脊椎动物的人工热休克元件表现更好。我们仔细确定了热休克的最佳条件，并定义了一个核心启动子用于未来的构建。最后，使用该系统，我们研究了过表达头部模式基因 Tc-orthodenticle1（Tc-otd1）的效果。我们发现 Tc-otd1 在躯干中抑制 Tc-wingless（Tc-wg），在头部中也有一定程度的抑制。
