报告使用 pORTMAGE 系统的重组工程的脱靶效应。
Reporting off-target effects of recombinant engineering using the pORTMAGE system.
机构信息
North Carolina Agricultural and Technical State University, USA.
North Carolina Agricultural and Technical State University, USA.
出版信息
J Microbiol Methods. 2023 Jan;204:106627. doi: 10.1016/j.mimet.2022.106627. Epub 2022 Nov 25.
Abstract
pORTMAGE recombineering is a simple technique for incorporation of novel point mutations into bacterial genomes that eliminates off-target effects. Here we inserted point mutations into the cusS gene from Escherichia coli, then, using Illumina sequencing, report genetic variants in all mutant strains. Several off-site mutations were found at high frequency. Low frequency mutations also show high heterogeneity. This means that it is essential for studies to report all off-target effects and acknowledge the effect that this may have on resultant phenotypes.
摘要
PORTMAGE 重组是一种将新的点突变引入细菌基因组的简单技术，可消除脱靶效应。在这里，我们将点突变插入大肠杆菌 cusS 基因中，然后使用 Illumina 测序报告所有突变株的遗传变异。发现了几个高频率的非靶标突变。低频突变也表现出高度的异质性。这意味着研究报告所有脱靶效应并承认这可能对最终表型产生影响是至关重要的。
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