González-Billault C, Avila J
Center of Molecular Biology Severo Ochoa, Universidad Autónoma de Madrid, Spain.
Histol Histopathol. 2000 Oct;15(4):1177-83. doi: 10.14670/HH-15.1177.
Protein function in vivo can be studied by deleting (knock-out) the gene that encodes it, and search for the consequences. This procedure involves different technologies, including recombinant DNA procedures, cell biology methods and histological and immunocytochemical analysis. In this work we have reviewed these procedures when they have been applied to ascertain the function of several microtubule-associated proteins. These proteins have been previously involved, through in vitro experiments, in having a role in the microtubule stabilization. Here, we will summarize the generation and characterization of different microtubule-associated protein knock-out mice. Special attention will be paid to MAP1B knock-out mice. Amongst the different MAPs knock-out mice these show the strongest phenotype, the most likely for being MAP1B, the MAP that is expressed earliest in neurogenesis. Molecular genetics could be considered as a valid and useful procedure to truly establish the in vivo functions of a protein, although it is necessary to be aware of possible artifacts such as the generation of some kinds of RNA alternative splicing. To avoid this the best strategy to be used must consider the deletion of the exon that contains the functional domains of the protein.
可通过删除（敲除）编码蛋白质的基因并探究其后果来研究蛋白质在体内的功能。该过程涉及多种技术，包括重组DNA技术、细胞生物学方法以及组织学和免疫细胞化学分析。在这项工作中，我们回顾了这些技术在用于确定几种微管相关蛋白功能时的应用情况。此前通过体外实验已表明这些蛋白在微管稳定中发挥作用。在此，我们将总结不同微管相关蛋白敲除小鼠的产生及特征。将特别关注MAP1B敲除小鼠。在不同的微管相关蛋白敲除小鼠中，这些小鼠表现出最强的表型，最有可能是MAP1B，即神经发生过程中最早表达的微管相关蛋白。分子遗传学可被视为真正确定蛋白质体内功能的有效且有用的方法，不过有必要留意可能出现的假象，比如某些种类RNA可变剪接的产生。为避免这种情况，所采用的最佳策略必须考虑删除包含蛋白质功能域的外显子。
