Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus-Reference-Cited by-同舟云学术

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

Published:2010-12 Issue:23 Volume:76 Page:7870-7877
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Fontaine Laetitia¹,Dandoy Damien¹,Boutry Céline¹,Delplace Brigitte¹,de Frahan Marie Henry¹,Fremaux Christophe²,Horvath Philippe²,Boyaval Patrick²,Hols Pascal¹

Affiliation:

1. Biochimie et Génétique Moléculaire Bactérienne, Institut des Sciences de la Vie, Université Catholique de Louvain, Place Croix du Sud 5/6, B-1348 Louvain-la-Neuve, Belgium

2. Danisco France SAS, BP 10, 86220 Dangé-Saint-Romain, France

Abstract

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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/AEM.01671-10

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