Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus

Laetitia Fontaine; Damien Dandoy; Céline Boutry; Brigitte Delplace; Marie Henry de Frahan; Christophe Fremaux; Philippe Horvath; Patrick Boyaval; Pascal Hols

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Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus

机译：基于自然转化的嗜热链球菌无标记靶向遗传修饰通用程序的开发

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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.

机译：建立了通用的自然转化方案，并成功将其应用于测试的19种嗜热链球菌菌株中的18种。该方案的效率使得能够使用体外扩增的诱变片段来执行大基因片段的缺失或插入。根据与突变相关的表型，可以分两步选择无标记的突变体，即使用适应性的Cre-loxP系统进行抗性标记插入和切除，也可以如此处所示，通过强大的阳性筛选程序一步选择，如此处所示的组氨酸原营养型。

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《Applied Microbiology》 |2010年第23期|共8页
作者
Laetitia Fontaine; Damien Dandoy; Céline Boutry; Brigitte Delplace; Marie Henry de Frahan; Christophe Fremaux; Philippe Horvath; Patrick Boyaval; Pascal Hols;
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1. Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus [J] . Laetitia Fontaine, Damien Dandoy, Céline Boutry, Applied and Environmental Microbiology . 2010,第23期

机译：基于自然转化的嗜热链球菌无标记靶向遗传修饰通用程序的开发
2. A versatile mini-mazF-cassette for marker-free targeted genetic modification in Bacillus subtilis [J] . Lin Zhiwei, Deng Bin, Jiao Zhihua, Journal of Microbiological Methods . 2013,第2期

机译：多功能mini-mazF盒式磁带，用于枯草芽孢杆菌的无标记靶向遗传修饰
3. Natural Genetic Transformation: a Novel Tool for Efficient Genetic Engineering of the Dairy Bacterium Streptococcus thermophilus [J] . Trinelise Blomqvist, Hilde Steinmoen, Leiv Sigve H?varstein Applied and Environmental Microbiology . 2006,第10期

机译：自然遗传转化：乳杆菌嗜热链球菌高效基因工程的新型工具。
4. On-line estimation of lactic acid concentration during batch fermentations of Streptococcus thermophilus based on pH measurement [C] . Peter Eduard, Rock Helmut 2012 12th International Conference on Control, Automation and Systems. . 2012

机译：基于pH测量的嗜热链球菌批量发酵过程中乳酸浓度的在线估算
5. Development of Peptide-Based Probes to Target Quorum Sensing in Streptococcus pneumoniae [D] . Koirala, Bimal. 2019

机译：基于肽的探针在链球菌肺炎链球菌靶向探测探针
6. Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus [O] . Laetitia Fontaine, Damien Dandoy, Céline Boutry, 2010

机译：基于自然转化的嗜热链球菌无标记靶向遗传修饰通用程序的开发
7. Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus▿ [O] . Fontaine, Laetitia, Dandoy, Damien, Boutry, Céline, 2010

机译：基于自然转化的嗜热链球菌无标记靶向遗传修饰通用方法的开发▿

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Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus

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