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BglBrick vectors and datasheets: A synthetic biology platform for gene expression

2011· article· en· 533 citations· W2107523606 on OpenAlex· 10.1186/1754-1611-5-12

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Opus teacher head0.027
GPT teacher head0.225
Teacher spread
0.198 · how far apart the two teachers sit on this one work
Validation status
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Abstract

BACKGROUND: As engineered biological systems become more complex, it is increasingly common to express multiple operons from different plasmids and inducible expression systems within a single host cell. Optimizing such systems often requires screening combinations of origins of replication, expression systems, and antibiotic markers. This procedure is hampered by a lack of quantitative data on how these components behave when more than one origin of replication or expression system are used simultaneously. Additionally, this process can be time consuming as it often requires the creation of new vectors or cloning into existing but disparate vectors. RESULTS: Here, we report the development and characterization of a library of expression vectors compatible with the BglBrick standard (BBF RFC 21). We have designed and constructed 96 BglBrick-compatible plasmids with a combination of replication origins, antibiotic resistance genes, and inducible promoters. These plasmids were characterized over a range of inducer concentrations, in the presence of non-cognate inducer molecules, and with several growth media, and their characteristics were documented in a standard format datasheet. A three plasmid system was used to investigate the impact of multiple origins of replication on plasmid copy number. CONCLUSIONS: The standardized collection of vectors presented here allows the user to rapidly construct and test the expression of genes with various combinations of promoter strength, inducible expression system, copy number, and antibiotic resistance. The quantitative datasheets created for these vectors will increase the predictability of gene expression, especially when multiple plasmids and inducers are utilized.

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The record

Venue
Journal of Biological Engineering
Topic
Bacterial Genetics and Biotechnology
Field
Biochemistry, Genetics and Molecular Biology
Canadian institutions
Funders
Lawrence Berkeley National LaboratoryBiological and Environmental ResearchNatural Sciences and Engineering Research Council of CanadaU.S. Department of EnergyOffice of ScienceNational Science Foundation
Keywords
PlasmidComputational biologyBiologyExpression vectorGeneOrigin of replicationGeneticsComputer scienceRecombinant DNA
Has abstract in OpenAlex
yes