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.
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。
相似文献
1
Development of a versatile procedure based on natural transformation for marker-free targeted genetic modification in Streptococcus thermophilus.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。
Appl Environ Microbiol. 2010 Dec;76(23):7870-7. doi: 10.1128/AEM.01671-10. Epub 2010 Oct 8.
2
Targeted mutagenesis of Burkholderia thailandensis and Burkholderia pseudomallei through natural transformation of PCR fragments.通过PCR片段的自然转化对泰国伯克霍尔德菌和类鼻疽伯克霍尔德菌进行靶向诱变。
Appl Environ Microbiol. 2008 May;74(10):2985-9. doi: 10.1128/AEM.00030-08. Epub 2008 Feb 29.
3
A food-grade site-directed mutagenesis system for Streptococcus thermophilus LMG 18311.用于嗜热链球菌 LMG 18311 的食品级定点突变系统。
Lett Appl Microbiol. 2010 Mar;50(3):314-9. doi: 10.1111/j.1472-765X.2009.02794.x. Epub 2009 Dec 24.
4
High-efficiency transformation of Streptococcus thermophilus using electroporation.利用电穿孔技术实现嗜热链球菌的高效转化。
J Sci Food Agric. 2021 Dec;101(15):6578-6585. doi: 10.1002/jsfa.11292. Epub 2021 May 14.
5
An efficient method to optimize Kluyveromyces lactis gene targeting.一种优化乳酸克鲁维酵母基因靶向的有效方法。
FEMS Yeast Res. 2011 Sep;11(6):509-13. doi: 10.1111/j.1567-1364.2011.00741.x. Epub 2011 Jun 29.
6
Transformation system for Hypocrea jecorina (Trichoderma reesei) that favors homologous integration and employs reusable bidirectionally selectable markers.棘孢木霉（里氏木霉）的转化系统，有利于同源整合，并采用可重复使用的双向选择标记。
Appl Environ Microbiol. 2011 Jan;77(1):114-21. doi: 10.1128/AEM.02100-10. Epub 2010 Nov 12.
7
Efficient method for generation of bacteriophage insensitive mutants of Streptococcus thermophilus yoghurt and mozzarella strains.生成嗜热链球菌酸奶和马苏里拉奶酪菌株噬菌体不敏感突变体的有效方法。
J Microbiol Methods. 2007 Jul;70(1):159-64. doi: 10.1016/j.mimet.2007.04.006. Epub 2007 Apr 24.
8
Burkholderia thailandensis: Genetic Manipulation.泰国伯克霍尔德菌：基因操作
Curr Protoc Microbiol. 2017 May 16;45:4C.2.1-4C.2.15. doi: 10.1002/cpmc.27.
9
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。
Biotechnol Lett. 2009 May;31(5):759-64. doi: 10.1007/s10529-009-9931-6. Epub 2009 Jan 27.
10
Recombineering in Using Direct Repeat-Mediated Cloning-Independent Markerless Mutagenesis (DR-CIMM).使用直接重复介导的无克隆标记诱变（DR-CIMM）进行重组工程。
Front Cell Infect Microbiol. 2017 May 23;7:202. doi: 10.3389/fcimb.2017.00202. eCollection 2017.
引用本文的文献
1
Selective degradation of phage RNAs by the Csm6 ribonuclease provides robust type III CRISPR immunity in Streptococcus thermophilus.Csm6 核糖核酸酶对噬菌体 RNA 的选择性降解为嗜热链球菌提供了强大的 III 型 CRISPR 免疫。
Nucleic Acids Res. 2024 Nov 11;52(20):12549-12564. doi: 10.1093/nar/gkae856.
2
Advances in Genetic Tools and Their Application in .基因工具的进展及其在……中的应用
Foods. 2023 Aug 19;12(16):3119. doi: 10.3390/foods12163119.
3
Bacteriophage-host interactions in Streptococcus thermophilus and their impact on co-evolutionary processes.热链球菌中噬菌体-宿主相互作用及其对共同进化过程的影响。
FEMS Microbiol Rev. 2023 Jul 5;47(4). doi: 10.1093/femsre/fuad032.
4
Investigation of CRISPR-Independent Phage Resistance Mechanisms Reveals a Role for FtsH in Phage Adsorption to Streptococcus thermophilus.CRISPR 独立噬菌体抗性机制的研究揭示了 FtsH 在噬菌体吸附到嗜热链球菌中的作用。
J Bacteriol. 2023 Jun 27;205(6):e0048222. doi: 10.1128/jb.00482-22. Epub 2023 May 31.
5
Development of a Tetracycline-Inducible System for Conditional Gene Expression in Lactococcus lactis and Streptococcus thermophilus.四环素诱导型系统在乳酸乳球菌和嗜热链球菌中的条件性基因表达的开发。
Microbiol Spectr. 2023 Jun 15;11(3):e0066823. doi: 10.1128/spectrum.00668-23. Epub 2023 May 16.
6
A Genome-Wide CRISPR Interference Screen Reveals an StkP-Mediated Connection between Cell Wall Integrity and Competence in Streptococcus salivarius.全基因组 CRISPR 干扰筛选揭示了唾液链球菌细胞壁完整性和感受态之间的 StkP 介导的连接。
mSystems. 2022 Dec 20;7(6):e0073522. doi: 10.1128/msystems.00735-22. Epub 2022 Nov 7.
7
Host-encoded, cell surface-associated exopolysaccharide required for adsorption and infection by lactococcal P335 phage subtypes.宿主编码的、细胞表面相关的胞外多糖，是乳球菌P335噬菌体亚型吸附和感染所必需的。
Front Microbiol. 2022 Oct 4;13:971166. doi: 10.3389/fmicb.2022.971166. eCollection 2022.
8
Competence shut-off by intracellular pheromone degradation in salivarius streptococci.唾液链球菌中细胞内信息素降解导致的失能。
PLoS Genet. 2022 May 25;18(5):e1010198. doi: 10.1371/journal.pgen.1010198. eCollection 2022 May.
9
Expanding natural transformation to improve beneficial lactic acid bacteria.扩大自然转化以改善有益乳酸菌。
FEMS Microbiol Rev. 2022 Jul 20;46(4). doi: 10.1093/femsre/fuac014.
10
The CovRS Environmental Sensor Directly Controls the ComRS Signaling System To Orchestrate Competence Bimodality in Salivarius Streptococci.柯维氏调控系统（CovRS 环境传感器）直接控制复合调控系统（ComRS 信号系统），从而协调唾液链球菌的兼性双态性。
mBio. 2022 Feb 22;13(1):e0312521. doi: 10.1128/mbio.03125-21. Epub 2022 Jan 4.
本文引用的文献
1
A food-grade site-directed mutagenesis system for Streptococcus thermophilus LMG 18311.用于嗜热链球菌 LMG 18311 的食品级定点突变系统。
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.
4
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.
5
Regulation of natural genetic transformation and acquisition of transforming DNA in Streptococcus pneumoniae.肺炎链球菌中自然遗传转化及转化DNA获取的调控
FEMS Microbiol Rev. 2009 May;33(3):627-42. doi: 10.1111/j.1574-6976.2009.00167.x.
6
Genome-scale model of Streptococcus thermophilus LMG18311 for metabolic comparison of lactic acid bacteria.用于乳酸菌代谢比较的嗜热链球菌LMG18311全基因组规模模型
Appl Environ Microbiol. 2009 Jun;75(11):3627-33. doi: 10.1128/AEM.00138-09. Epub 2009 Apr 3.
7
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。
Biotechnol Lett. 2009 May;31(5):759-64. doi: 10.1007/s10529-009-9931-6. Epub 2009 Jan 27.
8
Multifactorial experimental design for optimizing transformation: Electroporation of Streptococcus thermophilus.用于优化转化的多因素实验设计：嗜热链球菌的电穿孔法
Biotechnol Bioeng. 1994 Mar 15;43(6):490-6. doi: 10.1002/bit.260430609.
9
Markerless multiple-gene-deletion system for Streptococcus mutans.变形链球菌的无标记多基因缺失系统
Appl Environ Microbiol. 2008 Apr;74(7):2037-42. doi: 10.1128/AEM.02346-07. Epub 2008 Feb 8.
10
The inhibitory spectrum of thermophilin 9 from Streptococcus thermophilus LMD-9 depends on the production of multiple peptides and the activity of BlpG(St), a thiol-disulfide oxidase.嗜热链球菌LMD-9来源的嗜热菌素9的抑制谱取决于多种肽的产生以及硫醇-二硫键氧化酶BlpG(St)的活性。
Appl Environ Microbiol. 2008 Feb;74(4):1102-10. doi: 10.1128/AEM.02030-07. Epub 2007 Dec 21.
Zotero 插件