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Título

Generation of Food-Grade Recombinant Lactic Acid Bacterium Strains by Site-Specific Recombination

AutorMartín, M. Cruz ; Alonso, Juan Carlos ; Suárez Fernández, Juan Evaristo ; Álvarez González, Miguel Ángel
Palabras claveLactic acid bacteria
Recombinant microorganism
Food industry
Plasmid
Phage
Vector
Homologous recombination
Genetic engineering
Lactobacillus casei
Microorganism resistance
Fermentation starter
Virus
Lactobacillaceae
Bacteria
Fecha de publicaciónjun-2000
EditorAmerican Society for Microbiology
CitaciónApplied and Environmental Microbiology 66(6): 2599-2604 (2000)
ResumenThe construction of a delivery and clearing system for the generation of food-grade recombinant lactic acid bacterium strains, based on the use of an integrase (Int) and a resolvo-invertase (β-recombinase) and their respective target sites (attP-attB and six, respectively) is reported. The delivery system contains a heterologous replication origin and antibiotic resistance markers surrounded by two directly oriented six sites, a multiple cloning site where passenger DNA could be inserted (e.g., the cI gene of bacteriophage A2), the int gene, and the attP site of phage A2. The clearing system provides a plasmid-borne gene encoding β-recombinase. The nonreplicative vector-borne delivery system was transformed into Lactobacillus casei ATCC 393 and, by site-specific recombination, integrated as a single copy in an orientation- and Int-dependent manner into the attB site present in the genome of the host strain. The transfer of the clearing system into this strain, with the subsequent expression of the β-recombinase, led to site-specific DNA resolution of the non-food-grade DNA. These methods were validated by the construction of a stable food-grade L. casei ATCC 393-derived strain completely immune to phage A2 infection during milk fermentation.
URIhttp://hdl.handle.net/10261/5676
DOI10.1128/AEM.66.6.2599-2604.2000
ISSN0099-2240
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