黄亚佳,倪磊,金帆,杨光.基于高通量显微成像及分析技术的 DNA 重排研究[J].集成技术,2019,8(6):31-38
基于高通量显微成像及分析技术的 DNA 重排研究
Measurement of DNA Rearrangement by High-Throughput MicroscopyImaging and Analysis
  
DOI:
中文关键词:  高通量;显微成像和分析技术;DNA 重排;重复序列删除
英文关键词:high-throughput; microscopy imaging and analysis technology; DNA rearrangement; deletion of repeat sequences
基金项目:国家自然科学基金项目(31270150、21774039)
作者单位
黄亚佳 华中科技大学生命科学与技术学院 武汉 430074;中国科学院深圳先进技术研究院 深圳 518055 
倪磊 中国科学院深圳先进技术研究院 深圳 518055 
金帆 中国科学院深圳先进技术研究院 深圳 518055 
杨光 华中科技大学生命科学与技术学院 武汉 430074 
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中文摘要:
      直接的重复序列广泛地存在于真核和原核细胞基因组中,并且与多种疾病(如遗传性神经肌 肉神经退行性疾病等)相关,因此定量重复序列的删除变得非常重要。结合高通量显微成像和分析技术,该文设计了基于三色荧光报告系统的方法来定量重复序列删除的发生。结果显示,在铜绿假单胞菌中,重复序列的删除频率在 recA 基因缺失突变株中明显降低,而 RadA 蛋白和 UvrD 蛋白的缺失则会提高重复序列的删除频率,并且重复序列的删除与细菌的生长率和启动子等因素无关。该研究有助于加深对直接重复序列相关问题的理解,并为直接重复序列删除定量提供了新的方法。
英文摘要:
      Direct repeat sequences widely exist in eukaryotic and prokaryotic genomes, which involves in human diseases like hereditary neuromuscular neurodegenerative disease. Thus, it is of great importance to quantify the frequency of the deletion of direct repeat sequences. Using the high-throughput microscopy imaging and analysis technology, we designed a three-color fluorescent-based reporter system to quantify the frequency of the deletion of direct repeat sequences. We found that the frequency of the deletion of direct repeat sequences decreased in the recA mutant in P. aeruginosa, and the absence of RadA and UvrD increased the occurrence frequency of the deletion of direct repeat sequences, but the frequency of the deletion of direct repeat sequences was independent of the following factors: fitness or promoter variants etc. This work can be helpful to the further understanding of the deletion of direct repeat sequences, which also provide a novel method to quantify the deletion of direct repeat sequences.
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