Title
BglBricks: A flexible standard for biological part assembly
Date Issued
20 January 2010
Access level
open access
Resource Type
journal article
Author(s)
University of California
Publisher(s)
Springer Nature
Abstract
Background: Standard 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.Results: Here, 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.Conclusions: The 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. © 2010 Anderson et al; licensee BioMed Central Ltd.
Volume
4
Language
English
OCDE Knowledge area
Bioquímica, Biología molecular
Biología del desarrollo
Scopus EID
2-s2.0-77649305316
Source
Journal of Biological Engineering
ISSN of the container
1754-1611
DOI of the container
10.1186/1754-1611-4-1
Sponsor(s)
We thank Dr. Thomas Knight Jr. for inventing idempotent assembly and providing feedback on this new standard proposal. In addition, we thank the BioBricks Foundation for their work in supporting standardized biological parts. JCA was supported by a Damon Runyon Cancer Research Foundation Postdoctoral Fellowship. JED and GCW are supported by the UC Berkeley QB3 Institute and NSF Grant CBET-0756801. JCA, JED, ML, JAG, APA, and JDK are supported by the SynBERC NSF ERC [32]. JAG is supported by NSF Grant BES-0439124.
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