【摘要】 TALEs是在植物病原菌黄单胞杆菌(Xanthomonas)上发现的一类天然的DNA结合蛋白,其重复域已经成为模块化构建单元,用于人工DNA结合蛋白的构建。人工TALEs能够用于多种基因工程操作,包括转录调节和基因组编辑。为了在常规分子生物学实验室稳定、经济地构建TALE-TFs和TALENs,本文采用TALE Toolbox试剂盒,运用PCR扩增和"酶切-连接"克隆法构建人工TALEs。结果表明,该构建策略是一种快速高效构建TALEs的方法,能够在1周内完成TALEs构建。更多还原

【Abstract】 TALEs (Transcription activator like effectors) are a class of naturally DNA-binding proteins found in the plant pathogen Xanthomonas.Engineered repeat domains of TALE have been used as modular building units for construction of customized DNA-binding proteins.Artificial TALEs can be used for a wide variety of gene engineering applications , including transcriptional modulation and genome editing.The article describes process for reliable and affordable construction of TALE-TFs and TALENs by the combination of PCR amplification and′digestion-ligation′cloning based on the TALE Toolbox kit.The results show that the construction strategy is fast and efficient , and TALEs construction could be completed within 1week.更多还原