Advances of genetic engineering in streptococci and enterococci
Advances of genetic engineering in streptococci and enterococci
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DOI：
10.1111/1348-0421.13015
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发表时间：
2022-06
影响因子：
2.6
通讯作者：
Jun Kurushima;H. Tomita
中科院分区：
文献类型：
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作者：
Jun Kurushima;H. Tomita
在后基因组时代，反向基因工程是实验分子生物学不可或缺的方法，可以更深入地了解基因组特征和生物表型之间的主要关系。从技术上讲，基因工程是通过用设计的核苷酸序列替换靶基因组基因座来进行的，即所谓的定点诱变。为了通过同源重组人工操纵等位基因置换，研究人员已经改进了针对感兴趣的细菌物种进行优化的各种方法。在这里，我们回顾了广泛使用的基因工程技术，特别是链球菌和肠球菌，以及最近的进展，使更有效和灵活的操作。基于水平基因转移系统的基本生物学知识的合成生物学方法促进了基因工程的发展，例如天然接合转移，天然转化和CRISPR/Cas系统。因此，本文还描述了基因工程技术改进的基础分子生物学的基本见解。
In the post‐genome era, reverse genetic engineering is an indispensable methodology for experimental molecular biology to provide a deeper understanding of the principal relationship between genomic features and biological phenotypes. Technically, genetic engineering is carried out through allele replacement of a target genomic locus with a designed nucleotide sequence, so called site‐directed mutagenesis. To artificially manipulate allele replacement through homologous recombination, researchers have improved various methodologies that are optimized to the bacterial species of interest. Here, we review widely used genetic engineering technologies, particularly for streptococci and enterococci, and recent advances that enable more effective and flexible manipulation. The development of genetic engineering has been promoted by synthetic biology approaches based on basic biological knowledge of horizontal gene transfer systems, such as natural conjugative transfer, natural transformation, and the CRISPR/Cas system. Therefore, this review also describes basic insights into molecular biology that underlie improvements in genetic engineering technology.