dc.creator | Arce Rodríguez, Alejandro | es |
dc.creator | Benedetti, Ilaria | es |
dc.creator | Borrero de Acuña, José Manuel | es |
dc.creator | Silva Rocha, Rafael | es |
dc.creator | Lorenzo, Víctor de | es |
dc.date.accessioned | 2022-10-07T17:31:54Z | |
dc.date.available | 2022-10-07T17:31:54Z | |
dc.date.issued | 2021-09-17 | |
dc.identifier.citation | Arce Rodríguez, A., Benedetti, I., Borrero de Acuña, J.M., Silva Rocha, R. y Lorenzo, V.d. (2021). Standardization of inducer-activated broad host range expression modules: debugging and refactoring an alkane-responsive AlkS/PalkB device. Synthetic Biology, 6 (1), ysab030. https://doi.org/10.1093/synbio/ysab030. | |
dc.identifier.issn | 2397-7000 | es |
dc.identifier.uri | https://hdl.handle.net/11441/137738 | |
dc.description.abstract | Although inducible heterologous expression systems have been available since the birth of recombinant DNA technology, the diversity
of genetic devices and their coming together in the corresponding vectors often result in a lack of reproducibility and interoperability. In
an effort to increase the predictability of expression of genes of interest in a variety of possible bacterial hosts, we propose a composition
standard for debugging and reassembling all regulatory parts that participate in the performance of such devices. As a case study, we
addressed the n-octane and dicyclopropyl ketone-inducible PalkB promoter of the alkane biodegradation pOCT plasmid of Pseudomonas
putida. The standardized expression module included an edited alkS transcription factor divergently expressed and separated from
PalkB by a synthetic buffer segment. The DNA sequence of the alkS gene was modified to alleviate the catabolite repression exerted
by several carbon and nitrogen sources through the Crc/Hfq complex of some hosts. The PalkB promoter and the alkS variants were
then formatted as Standard European Vector Architecture cargoes, and their activity in P. putida was quantified with fluorescent and
luminescent reporters. Despite considerable editing of the DNA sequences involved, the thereby refactored module basically kept the
functioning parameters of the original configuration. The same qualities were inspected when the system was passed to Escherichia
coli and Pseudomonas aeruginosa. We argue that the application of the compositional standard thereby implemented in the AlkS/PalkB
module to other promoter/regulator pairs will enable more complex genetic programming in non-model bacteria. | es |
dc.description.sponsorship | MADONNA H2020-FET OPEN-RIA-2017-1-766975 | es |
dc.description.sponsorship | BioRoboost H2020-NMBP-BIO-CSA 2018-820699 | es |
dc.description.sponsorship | SynBio4Flav H2020-NMBP-TR-IND/H2020-NMBP BIO-2018-814650 | es |
dc.description.sponsorship | MIX-UP H2020-BIO-CN-2019-870294 | es |
dc.format | application/pdf | es |
dc.format.extent | 11 p. | es |
dc.language.iso | eng | es |
dc.publisher | Oxford University Press | es |
dc.relation.ispartof | Synthetic Biology, 6 (1), ysab030. | |
dc.rights | Attribution-NonCommercial-NoDerivatives 4.0 Internacional | * |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | * |
dc.subject | SEVA | es |
dc.subject | AlkS | es |
dc.subject | Pseudomonas | es |
dc.subject | Crc | es |
dc.subject | Hfq | es |
dc.subject | cytometry | es |
dc.subject | noise | es |
dc.title | Standardization of inducer-activated broad host range expression modules: debugging and refactoring an alkane-responsive AlkS/PalkB device | es |
dc.type | info:eu-repo/semantics/article | es |
dcterms.identifier | https://ror.org/03yxnpp24 | |
dc.type.version | info:eu-repo/semantics/publishedVersion | es |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | es |
dc.contributor.affiliation | Universidad de Sevilla. Departamento de Microbiología | es |
dc.relation.publisherversion | https://watermark.silverchair.com/ysab030.pdf?token=AQECAHi208BE49Ooan9kkhW_Ercy7Dm3ZL_9Cf3qfKAc485ysgAAAuEwggLdBgkqhkiG9w0BBwagggLOMIICygIBADCCAsMGCSqGSIb3DQEHATAeBglghkgBZQMEAS4wEQQMX3TLQU4OWkmHC2XQAgEQgIIClCvQ7kWRo3VAIpqp3zpFEblCA17JK7YEiM39SVK66dQrNyF9wSQxrcWckQrP08L3QICZvj_Dg5uRqTWPfijuZc8DPZZbq7uvxLjhyvVaTTpQmvUcyLAXYCgi6DWzELBRfyFZ8IZOustPnPGNuv6imJOonOxveTP626orXJ8hSvCluZswO7sopff_5rHknUTJkt6bJiw5QuS62kAG7S4h4YRcLuTPm10xCXOkfZZK1nNrUWACUcnuqlIjPufjKaB2oTkjH0ennt3Kcg3YKiKmuA16D0n-QZDAJ7oRX5OgKjcMD4CUrisWgDtfT7n63jRXyTCWowPh76gYKH7ZJ3UptGguwqxGoa4Dd5sUCo4zp2bkHtL7-H7pwZs720tQIQ7o0a81IvuYjc2h6TaogbjspQO0ZOQzoxZV6PeaW5iRV7-eyS0KEjLOgNvwKNj4uWwJBAaXKpJ3m1p39CUq_s1EZFccqvwqEa0cgubtLIE8mYWnjaTor2rYirCOiRJrR5Uckpx2p8bxmTuvJz5rkYgW747zGt1ZBNaF2qf-Alj65CpPdqOOZ-O0G-KNzsydsrnsPiSG2s21KCGQh1CkPhYP8WmlvJKMLBCXd353Cv-STIOZ8Sb_Dn5PsrTLMQjOAL7fEGajwTdKZOYJPKLNubXS6KFz2rVAA5_IK_hslIAjJlGwwyOZuTMXiAE0r_lekONLO8bZ-a_4Tq7sk9-5Z5jarbAAiJDgoKhmSGqAKgN2rkKoC6kjV1fMBBob_jJgz5nDVD8tQNXLmcjbPcrwcFwa9yI__07RADLJosDD6s1vEh7tl2ietMvzSqg3xt16RGOu65wJDk7RKInt4ALBRjUA2n8hFnwHDQ5c_VBQKiDBJkh51uuaTA | es |
dc.identifier.doi | 10.1093/synbio/ysab030 | es |
dc.journaltitle | Synthetic Biology | es |
dc.publication.volumen | 6 | es |
dc.publication.issue | 1 | es |
dc.publication.initialPage | ysab030 | |
dc.contributor.funder | Ministerio de Ciencia e Innovación (MICIN). España | es |