| dc.contributor |
European Commission |
|
| dc.creator |
Martín, M. Cruz |
|
| dc.creator |
Alonso, Juan Carlos |
|
| dc.creator |
Suárez Fernández, Juan Evaristo |
|
| dc.creator |
Álvarez González, Miguel Ángel |
|
| dc.date |
2008-07-08T12:13:25Z |
|
| dc.date |
2008-07-08T12:13:25Z |
|
| dc.date |
2000-06 |
|
| dc.date.accessioned |
2017-01-31T02:42:55Z |
|
| dc.date.available |
2017-01-31T02:42:55Z |
|
| dc.identifier |
Applied and Environmental Microbiology 66(6): 2599-2604 (2000) |
|
| dc.identifier |
0099-2240 |
|
| dc.identifier |
http://hdl.handle.net/10261/5676 |
|
| dc.identifier |
10.1128/AEM.66.6.2599-2604.2000 |
|
| dc.identifier.uri |
http://dspace.mediu.edu.my:8181/xmlui/handle/10261/5676 |
|
| dc.description |
The 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. |
|
| dc.description |
This work was supported by grant BIO4-CT96-0402 to J.E.S. and grants PB 96-0817 and BIO4-CT98-0250 to J.C.A. |
|
| dc.description |
Peer reviewed |
|
| dc.format |
22195 bytes |
|
| dc.format |
application/pdf |
|
| dc.language |
eng |
|
| dc.publisher |
American Society for Microbiology |
|
| dc.rights |
closedAccess |
|
| dc.subject |
Lactic acid bacteria |
|
| dc.subject |
Recombinant microorganism |
|
| dc.subject |
Food industry |
|
| dc.subject |
Plasmid |
|
| dc.subject |
Phage |
|
| dc.subject |
Vector |
|
| dc.subject |
Homologous recombination |
|
| dc.subject |
Genetic engineering |
|
| dc.subject |
Lactobacillus casei |
|
| dc.subject |
Microorganism resistance |
|
| dc.subject |
Fermentation starter |
|
| dc.subject |
Virus |
|
| dc.subject |
Lactobacillaceae |
|
| dc.subject |
Bacteria |
|
| dc.title |
Generation of Food-Grade Recombinant Lactic Acid Bacterium Strains by Site-Specific Recombination |
|
| dc.type |
Artículo |
|