Registro completo de metadatos
| Campo DC | Valor | Lengua/Idioma |
|---|---|---|
| dc.creator | Hernández Tamayo, Rogelio | - |
| dc.creator | Torres Tejerizo, Gonzalo Arturo | - |
| dc.creator | Brom, Susana | - |
| dc.creator | Romero, David | - |
| dc.date | 2018-06-11T20:58:44Z | - |
| dc.date | 2018-06-11T20:58:44Z | - |
| dc.date | 2016-06 | - |
| dc.date | 2018-06-08T14:26:44Z | - |
| dc.date.accessioned | 2019-04-29T15:28:08Z | - |
| dc.date.available | 2019-04-29T15:28:08Z | - |
| dc.date.issued | 2018-06-11T20:58:44Z | - |
| dc.date.issued | 2018-06-11T20:58:44Z | - |
| dc.date.issued | 2016-06 | - |
| dc.date.issued | 2018-06-08T14:26:44Z | - |
| dc.identifier | Hernández Tamayo, Rogelio; Torres Tejerizo, Gonzalo Arturo; Brom, Susana; Romero, David; Site-specific bacterial chromosome engineering mediated by IntA integrase from Rhizobium etli; BioMed Central; BMC Microbiology; 16; 1; 6-2016; 1-9 | - |
| dc.identifier | 1471-2180 | - |
| dc.identifier | http://hdl.handle.net/11336/48192 | - |
| dc.identifier | CONICET Digital | - |
| dc.identifier | CONICET | - |
| dc.identifier.uri | http://rodna.bn.gov.ar:8080/jspui/handle/bnmm/294683 | - |
| dc.description | Background: The bacterial chromosome may be used to stably maintain foreign DNA in the mega-base range. Integration into the chromosome circumvents issues such as plasmid replication, stability, incompatibility, and copy number variance. The site-specific integrase IntA from Rhizobium etli CFN42 catalyzes a direct recombination between two specific DNA sites: attA and attD (23 bp). This recombination is stable. The aim of this work was to develop a R. etli derivative that may be used as recipient for the integration of foreign DNA in the chromosome, adapting the IntA catalyzed site-specific recombination system. Results: To fulfill our aim, we designed a Rhizobium etli CFN42 derivative, containing a “landing pad” (LP) integrated into the chromosome. The LP sector consists of a green fluorescent protein gene under the control of the lacZ promoter and a spectinomycin resistance gene. Between the lacZ promoter and the GFP gene we inserted an IntA attachment site, which does not affect transcription from the lac promoter. Also, a mobilizable donor vector was generated, containing an attA site and a kanamycin resistance gene; to facilitate insertion of foreign DNA, this vector also contains a multicloning site. There are no promoters flanking the multicloning site. A biparental mating protocol was used to transfer the donor vector into the landing pad strain; insertion of the donor vector into the landing pad sector via IntA-mediated attA X attA recombination thereby interrupted the expression of the green fluorescent protein, generating site-specific cointegrants. Cointegrants were easily recognized by screening for antibiotic sensitivity and lack of GFP expression, and were obtained with an efficiency of 6.18 %. Conclusions: Integration of foreign DNA in Rhizobium, lacking any similarity with the genome, can be easily achieved by IntA-mediated recombination. This protocol contains the mating and selection procedures for creating and isolating integrants. | - |
| dc.description | Fil: Hernández Tamayo, Rogelio. Universidad Nacional Autónoma de México; México | - |
| dc.description | Fil: Torres Tejerizo, Gonzalo Arturo. Universidad Nacional Autónoma de México; México. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina | - |
| dc.description | Fil: Brom, Susana. Universidad Nacional Autónoma de México; México | - |
| dc.description | Fil: Romero, David. Universidad Nacional Autónoma de México; México | - |
| dc.format | application/pdf | - |
| dc.format | application/pdf | - |
| dc.language | eng | - |
| dc.publisher | BioMed Central | - |
| dc.relation | info:eu-repo/semantics/altIdentifier/doi/https://dx.doi.org/10.1186/s12866-016-0755-y | - |
| dc.relation | info:eu-repo/semantics/altIdentifier/url/https://bmcmicrobiol.biomedcentral.com/articles/10.1186/s12866-016-0755-y | - |
| dc.rights | info:eu-repo/semantics/openAccess | - |
| dc.rights | https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ | - |
| dc.source | reponame:CONICET Digital (CONICET) | - |
| dc.source | instname:Consejo Nacional de Investigaciones Científicas y Técnicas | - |
| dc.source | instacron:CONICET | - |
| dc.subject | Chromosomal integration | - |
| dc.subject | Site-specific recombination | - |
| dc.subject | Tyrosine recombinase | - |
| dc.subject | Otras Ciencias Biológicas | - |
| dc.subject | Ciencias Biológicas | - |
| dc.subject | CIENCIAS NATURALES Y EXACTAS | - |
| dc.title | Site-specific bacterial chromosome engineering mediated by IntA integrase from Rhizobium etli | - |
| dc.type | info:eu-repo/semantics/article | - |
| dc.type | info:eu-repo/semantics/publishedVersion | - |
| dc.type | info:ar-repo/semantics/articulo | - |
| Aparece en las colecciones: | CONICET | |
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