BglBricks: A flexible standard for biological part assembly

被引：238

作者：

Anderson J.C. ^{[1
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Dueber J.E. ^{[1
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Leguia M. ^{[1
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Wu G.C. ^{[1
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Arkin A.P. ^{[1
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Keasling J.D. ^{[1
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机构：

[1] Department of Bioengineering, University of California, Berkeley

[2] California Institute for Quantitative Biological Research, University of California, Berkeley

[3] Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley

[4] Synthetic Biology Engineering Research Center, University of California, Berkeley

[5] Joint BioEnergy Institute, Emeryville

[6] Department of Chemical Engineering, University of California, Berkeley

来源：

Journal of Biological Engineering | / 4卷 / 1期

基金：

美国国家科学基金会;

关键词：

Synthetic Biology; Composite Part; Helper Plasmid; Assembly Reaction; Antibiotic Resistance Marker;

D O I：

10.1186/1754-1611-4-1

中图分类号：

学科分类号：

摘要：

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.

引用

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