BglBricks: A flexible standard for biological part assembly

By JChristopher Anderson, John E. Dueber, Mariana Leguia, Gabriel C. Wu, Jonathan A. Goler, Adam P. Arkin, Jay D. Keasling. Journal of Biological Engineering, 2010

454 citations recorded by the source corpus.

BackgroundStandard biological parts, such as BioBricks™ parts, provide the foundation for a new engineering discipline that enables the design and construction of synthetic biological systems with a variety of applications in bioenergy, new materials, therapeutics, and environmental remediation. Although the original BioBricks™ assembly standard has found widespread use, it has several shortcomings that limit its range of potential applications. In particular, the system is not suitable for the construction of protein fusions due to an unfavorable scar sequence that encodes an in-frame stop codon.ResultsHere, we present a similar but new composition standard, called BglBricks, that addresses the scar translation issue associated with the original standard. The new system employs BglII and BamHI restriction enzymes, robust cutters with an extensive history of use, and results in a 6-nucleotide scar sequence encoding glycine-serine, an innocuous peptide linker in most protein fusion applications. We demonstrate the utility of the new standard in three distinct applications, including the construction of constitutively active gene expression devices with a wide range of expression profiles, the construction of chimeric, multi-domain protein fusions, and the targeted integration of functional DNA sequences into specific loci of the E. coli genome.ConclusionsThe BglBrick standard provides a new, more flexible platform from which to generate standard biological parts and automate DNA assembly. Work on BglBrick assembly reactions, as well as on the development of automation and bioinformatics tools, is currently underway. These tools will provide a foundation from which to transform genetic engineering from a technically intensive art into a purely design-based discipline.

Subfields: Mechanical Engineering, Molecular Biology, Radiology, Nuclear Medicine and Imaging

View Paper: https://www.semanticscholar.org/paper/4d9c45e437b15dedac830387a9cd30213e4a5060

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In Molecular Biology

Citations in the corpus

A Modular Cloning System for Standardized Assembly of Multigene Constructs (2011, background): “Since the first BioBrick standard, various standards and assembly protocols have been developed in order to optimize the sequence junctions between parts or make cloning more efficient [11,12].”
The Standard European Vector Architecture (SEVA): a coherent platform for the analysis and deployment of complex prokaryotic phenotypes (2012, methodology): “A variant of the BioBrick approach is the BglBrick approach (23), which exploits the loss of BlgII and BamHI sites when DNA segments that are cleaved with either enzyme are ligated to each other in a fashion that allows for regeneration of a usable BlgII sequence after each round of insertion.”
Identification and microbial production of a terpene-based advanced biofuel (2011, methodology): “Mevalonate pathway vectors pBb plasmids were prepared as the BglBrick standard expression vector”