Harder Ben, Schomburg Adrian, Pflanz Ralf, Küstner Katharina, Gerlach Nina, Schuh Reinhard
Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany.
Biotechniques. 2008 May;44(6):765-72. doi: 10.2144/000112884.
Drosophila provides a powerful experimental system to analyze gene functions in a multi-cellular organism. Here we describe an in vivo method that interferes with the integrity of selected proteins through site-specific cleavage in Drosophila. The technique is based on the highly specific seven-amino-acid recognition site of the tobacco etch virus (TEV) protease. We established transgenic fly lines that direct TEV protease expression in various tissues without affecting fly viability. The insertion of the TEV protease recognition site in defined positions of target proteins mediates their sequence-specific cleavage after controlled TEV protease expression in the fly. Thereby, this technique is a powerful tool that allows the in vivo manipulation of selected proteins in a time- and tissue-specific manner.
果蝇为分析多细胞生物中的基因功能提供了一个强大的实验系统。在此，我们描述一种体内方法，该方法通过在果蝇中进行位点特异性切割来干扰选定蛋白质的完整性。该技术基于烟草蚀纹病毒（TEV）蛋白酶的高度特异性七氨基酸识别位点。我们建立了转基因果蝇品系，可在不影响果蝇生存能力的情况下在各种组织中指导TEV蛋白酶的表达。在果蝇中受控表达TEV蛋白酶后，将TEV蛋白酶识别位点插入靶蛋白的特定位置可介导其序列特异性切割。因此，该技术是一种强大的工具，能够以时间和组织特异性方式在体内操纵选定的蛋白质。
