The protein trap is a powerful tool for genetic and biochemical studies of gene function in the animal kingdom. Although the original protein trap was developed for flies, it can be easily adapted to other multicellular organisms, both known models and ones with an unsequenced genome. The protein trap has been successfully applied to the fruit fly, crustaceans Parhyale hawaiensis, zebrafish, and insect and animal cell cultures. This approach is based on the integration into genes of an artificial exon that carries DNA encoding a fluorescent marker, standardized immunoepitopes, an integrase docking site, and splice acceptor and donor sites. The protein trap for cell cultures additionally contains an antibiotic resistance gene, which facilitates the selection of trapped clones. Resulting chimeric tagged mRNAs can be interfered by dsRNA against GFP (iGFPi-in vivo GFP interference), or the chimeric proteins can be efficiently knocked down by deGradFP technology. Both RNA and protein knockdowns produce a strong loss of function phenotype in tagged cells. The fluorescent and protein affinity tags can be used for tagged protein localisation within the cell and for identifying their binding partners in their native complexes. Insertion into protein trap integrase docking sites allows the replacement of trap contents by any new constructs, including other markers, cell toxins, stop-codons, and binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS, that reliably reflect endogenous gene expression. A distinctive feature of the protein trap approach is that all manipulations with a gene or its product occur only in the endogenous locus, which cannot be achieved by any other method.

中文翻译：

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蛋白质陷阱：遗传学家的新瑞士军刀？

蛋白质陷阱是动物界中基因功能的遗传和生化研究的有力工具。尽管最初的蛋白质捕获器是为果蝇开发的，但它可以轻松地适应其他多细胞生物，包括已知模型和基因组未测序的模型。该蛋白捕集器已成功应用于果蝇，甲壳类夏威夷Parhyale，斑马鱼以及昆虫和动物细胞培养。该方法基于将带有编码荧光标记，标准化免疫表位，整合酶停靠位点以及剪接受体和供体位点的DNA的人工外显子整合到基因中。用于细胞培养的蛋白质捕获器还包含一个抗生素抗性基因，这有助于选择捕获的克隆。dsRNA可以干扰GFP产生的嵌合标记的mRNA（iGFPi-体内GFP干扰），或者可以通过deGradFP技术有效地敲除嵌合蛋白。RNA和蛋白敲低都在标记细胞中产生功能表型的强烈丧失。荧光和蛋白质亲和标签可用于标记的蛋白质在细胞内的定位，并用于识别其天然复合物中的结合伴侣。插入蛋白质陷阱整合酶停靠位点后，可以用任何新构建体替换陷阱内容，包括其他标记，细胞毒素，终止密码子和诸如GAL4 / UAS，LexA / LexAop和QF / QUAS之类的二元表达系统内源基因表达。