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pBLAM1-x: standardized transposon tools for high-throughput screening

The engineering of pre-defined functions in living cells requires increasingly accurate tools as synthetic biology efforts become more ambitious. Moreover, the characterization of the phenotypic performance of genetic constructs demands meticulous measurements and extensive data acquisition for the...

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Autores principales: Alejaldre, Lorea, Anhel, Ana-Mariya, Goñi-Moreno, Ángel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10306358/
https://www.ncbi.nlm.nih.gov/pubmed/37388964
http://dx.doi.org/10.1093/synbio/ysad012
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author Alejaldre, Lorea
Anhel, Ana-Mariya
Goñi-Moreno, Ángel
author_facet Alejaldre, Lorea
Anhel, Ana-Mariya
Goñi-Moreno, Ángel
author_sort Alejaldre, Lorea
collection PubMed
description The engineering of pre-defined functions in living cells requires increasingly accurate tools as synthetic biology efforts become more ambitious. Moreover, the characterization of the phenotypic performance of genetic constructs demands meticulous measurements and extensive data acquisition for the sake of feeding mathematical models and matching predictions along the design-build-test lifecycle. Here, we developed a genetic tool that eases high-throughput transposon insertion sequencing (TnSeq): the pBLAM1-x plasmid vectors carrying the Himar1 Mariner transposase system. These plasmids were derived from the mini-Tn5 transposon vector pBAMD1-2 and built following modular criteria of the Standard European Vector Architecture (SEVA) format. To showcase their function, we analyzed sequencing results of 60 clones of the soil bacterium Pseudomonas putida KT2440. The new pBLAM1-x tool has already been included in the latest SEVA database release, and here we describe its performance using laboratory automation workflows. Graphical Abstract [Image: see text]
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spelling pubmed-103063582023-06-29 pBLAM1-x: standardized transposon tools for high-throughput screening Alejaldre, Lorea Anhel, Ana-Mariya Goñi-Moreno, Ángel Synth Biol (Oxf) Short Communication The engineering of pre-defined functions in living cells requires increasingly accurate tools as synthetic biology efforts become more ambitious. Moreover, the characterization of the phenotypic performance of genetic constructs demands meticulous measurements and extensive data acquisition for the sake of feeding mathematical models and matching predictions along the design-build-test lifecycle. Here, we developed a genetic tool that eases high-throughput transposon insertion sequencing (TnSeq): the pBLAM1-x plasmid vectors carrying the Himar1 Mariner transposase system. These plasmids were derived from the mini-Tn5 transposon vector pBAMD1-2 and built following modular criteria of the Standard European Vector Architecture (SEVA) format. To showcase their function, we analyzed sequencing results of 60 clones of the soil bacterium Pseudomonas putida KT2440. The new pBLAM1-x tool has already been included in the latest SEVA database release, and here we describe its performance using laboratory automation workflows. Graphical Abstract [Image: see text] Oxford University Press 2023-06-21 /pmc/articles/PMC10306358/ /pubmed/37388964 http://dx.doi.org/10.1093/synbio/ysad012 Text en © The Author(s) 2023. Published by Oxford University Press. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Short Communication
Alejaldre, Lorea
Anhel, Ana-Mariya
Goñi-Moreno, Ángel
pBLAM1-x: standardized transposon tools for high-throughput screening
title pBLAM1-x: standardized transposon tools for high-throughput screening
title_full pBLAM1-x: standardized transposon tools for high-throughput screening
title_fullStr pBLAM1-x: standardized transposon tools for high-throughput screening
title_full_unstemmed pBLAM1-x: standardized transposon tools for high-throughput screening
title_short pBLAM1-x: standardized transposon tools for high-throughput screening
title_sort pblam1-x: standardized transposon tools for high-throughput screening
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10306358/
https://www.ncbi.nlm.nih.gov/pubmed/37388964
http://dx.doi.org/10.1093/synbio/ysad012
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