Abstract：
The technical challenge in proving that a given expressed pseudogene is in fact translated into a functional protein is specificity. To circumvent this challenge, one approach is to use PCR in order to generate a series of clones that allow one to exogenously express the pseudogenic protein of interest, either native or fused to a tag, which can facilitate purification, detection, and complementation in both bacterial and mammalian cells. This approach allows an assessment of whether a putative pseudogenic protein possesses enzymatic activity, to identify its subcellular localization and to test its capacity to complement the parental homolog. An alternative approach is to detect the endogenous protein using targeted proteomics analysis and to assess the full range of endogenous RNA isoforms, in order to consider additional coding and noncoding RNA functionality.
摘要：
证明给定表达的假基因实际上被翻译成功能蛋白的技术挑战是特异性。为了规避这一挑战，一种方法是使用PCR来产生一系列克隆，这些克隆允许外源表达感兴趣的假蛋白，本地或融合到一个标签，这可以促进净化，检测,以及细菌和哺乳动物细胞中的互补。这种方法可以评估假定的假蛋白是否具有酶活性，以鉴定其亚细胞定位并测试其补充亲本同源物的能力。另一种方法是使用靶向蛋白质组学分析来检测内源性蛋白质，并评估内源性RNA亚型的全部范围。以考虑额外的编码和非编码RNA功能。
