反选标记法在微生物基因敲除中的应用

Application of the Efficient Counter-selection Method in Gene Deletion of Microorganism

随着后基因组时代的到来,简单高效的基因编辑工具和方法在研究微生物的基因表达调控、工程菌株的遗传改造等方面发挥了巨大作用。反选标记法是基于同源重组原理进行基因敲除的一种便捷高效的方法,该方法通过选择性的遗传标记的丢失和回补,实现对靶基因的敲除或目的基因的插入等分子遗传操作。这种选择性的遗传标记一般不依赖于传统的抗生素筛选标签,且同一反选标记能够反复使用,可连续进行多轮遗传操作,实现对多个基因靶位点的分子编辑。对近年来反选标记基因的种类、作用原理及在微生物基因敲除实验中的应用进行综述,以期为高效、可靠的分子编辑方法的研究提供参考。

With the coming of the post genome era,simple and efficient gene editing tools and methods have played a critical role in studying the regulation of gene expression and the genetic modification of engineered strains.The counter-selection methods are much convenient and efficient for gene knockout,which are based on the principle of homologous recombination.Knockout or insert of one target gene is achieved by the loss or supplement of one selective genetic marker,which generally does not rely on the traditional antibiotics.To achieve genetic manipulation of more target genes,the same selected marker can be used repeatedly over the progress of successive manipulation rounds.Because of its versatility,simplicity,efficacy,and wide range of applications,the counter-selective system has been applied in many fields of research,such as biotechnology,genetic engineering,and fundamental and applied biology.In recent years,the types,mechanisms and applications of counter-selection marker genes in microbial gene knockout experiments are reviewed.This genome-editing technique has a valuable advantage that particularly attracts the scientist’s mind,as it allows genome editing in multiple biological systems,and supports great opportunities for the future development of microbial science and rapid remodeling of genes.