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开发一种基于自然转化的通用程序，用于在嗜热链球菌中进行无标记靶向基因修饰。

Development of a versatile procedure based on natural transformation for marker-free targeted genetic modification in Streptococcus thermophilus.

机构信息

Institut des Sciences de la Vie, Université Catholique de Louvain, Place Croix du Sud 5, Box 6, B-1348 Louvain-la-Neuve, Belgium.

出版信息

Appl Environ Microbiol. 2010 Dec;76(23):7870-7. doi: 10.1128/AEM.01671-10. Epub 2010 Oct 8.

DOI:10.1128/AEM.01671-10
PMID:20935129
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2988589/
Abstract

A versatile natural transformation protocol was established for and successfully applied to 18 of the 19 Streptococcus thermophilus strains tested. The efficiency of the protocol enables the use of in vitro-amplified mutagenesis fragments to perform deletion or insertion of large genetic fragments. Depending on the phenotype linked to the mutation, markerless mutants can be selected either in two steps, i.e., resistance marker insertion and excision using an adapted Cre-loxP system, or in one step using a powerful positive screening procedure as illustrated here for histidine prototrophy.

摘要

建立了一种通用的自然转化方案，并成功应用于 19 株嗜热链球菌中的 18 株。该方案的效率使得可以使用体外扩增的诱变片段来进行大片段基因的缺失或插入。根据与突变相关的表型，可以使用两种方法选择无标记突变体，即使用改良的 Cre-loxP 系统插入和切除抗性标记，或者使用这里所示的强大的阳性筛选程序一步选择，如组氨酸营养缺陷型。

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Lett Appl Microbiol. 2010 Mar;50(3):314-9. doi: 10.1111/j.1472-765X.2009.02794.x. Epub 2009 Dec 24.
2
A novel pheromone quorum-sensing system controls the development of natural competence in Streptococcus thermophilus and Streptococcus salivarius.一种新型的信息素群体感应系统控制了嗜热链球菌和唾液链球菌自然感受态的发育。
J Bacteriol. 2010 Mar;192(5):1444-54. doi: 10.1128/JB.01251-09. Epub 2009 Dec 18.
3
A self-deleting Cre-lox-ermAM cassette, Cheshire, for marker-less gene deletion in Streptococcus pneumoniae.一种自我删除的 Cre-lox-ermAM 盒式元件 Cheshire，用于肺炎链球菌中的无标记基因缺失。
J Microbiol Methods. 2009 Dec;79(3):353-7. doi: 10.1016/j.mimet.2009.10.007. Epub 2009 Oct 20.
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The oligopeptide transport system is essential for the development of natural competence in Streptococcus thermophilus strain LMD-9.寡肽转运系统对于嗜热链球菌LMD-9菌株自然感受态的发展至关重要。
J Bacteriol. 2009 Jul;191(14):4647-55. doi: 10.1128/JB.00257-09. Epub 2009 May 15.
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Insertion of a heterologous gene construct into a non-functional ORF of the Streptococcus thermophilus chromosome.将异源基因构建体插入嗜热链球菌染色体的无功能开放阅读框中。
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8
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