Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo, 062-8517, Japan.
Plant Sci. 2021 Nov;312:111023. doi: 10.1016/j.plantsci.2021.111023. Epub 2021 Aug 15.
Genome-editing technologies are widely used to characterize gene functions and improve the features of agricultural plants. Although sequence analysis of gene editing target DNA is the most reliable method of screening gene-edited plants, the current DNA sequence analysis methods are time consuming and labor intensive because they include genomic DNA and polymerase chain reaction (PCR) product purification. In this study, seven methods were performed for sequence analysis of plant genomic DNA with and/or without genomic DNA and PCR product purification. Consequently, good-quality sequencing chromatograms were obtained using all methods. Results showed that the partial genomic DNA sequence of Nicotiana benthamiana and Arabidopsis thaliana could be sufficiently analyzed without plant genomic DNA and PCR product purification. Furthermore, screening of gene-edited N. benthamiana was successful using the present methods. Therefore, the tested methods could reduce the time, simplify the workflow of plant gene analysis, and help in screening gene-edited plants.
基因编辑技术广泛用于研究基因功能和改良农作物的特性。尽管对基因编辑靶标 DNA 的序列分析是筛选基因编辑植物最可靠的方法，但由于包括基因组 DNA 和聚合酶链式反应（PCR）产物纯化在内的当前 DNA 序列分析方法耗时耗力。本研究对有和/或无基因组 DNA 和 PCR 产物纯化的植物基因组 DNA 进行了 7 种方法的序列分析。结果表明，无需对植物基因组 DNA 和 PCR 产物进行纯化，即可充分分析本氏烟和拟南芥的部分基因组 DNA 序列。此外，本研究方法还成功筛选到基因编辑本氏烟。因此，这些方法可减少时间，简化植物基因分析的工作流程，有助于筛选基因编辑植物。
