Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria
Abstract
Molecular genetics is indispensable for interrogating the physiology of bacteria. However, the development of a functional genetic system for any given bacterium can be time-consuming. Here, we present a streamlined approach for identifying an effective transposon mutagenesis system for a new bacterium. Our strategy first involves the construction of hundreds of different transposon vector variants, which we term a “magic pool.” The efficacy of each vector in a magic pool is monitored in parallel using a unique DNA barcode that is introduced into each vector design. Using archived DNA “parts,” we next reassemble an effective vector for making a whole-genome transposon mutant library that is suitable for large-scale interrogation of gene function using competitive growth assays. Here, we demonstrate the utility of the magic pool system to make mutant libraries in five genera of bacteria.
- Authors:
-
[1];
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Div.
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth and Environmental Sciences Area
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Div.; Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1434009
- Alternate Identifier(s):
- OSTI ID: 1582038
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- mSystems
- Additional Journal Information:
- Journal Volume: 3; Journal Issue: 1; Journal ID: ISSN 2379-5077
- Publisher:
- American Society for Microbiology
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; genomics; transposons
Citation Formats
Liu, Hualan, Price, Morgan N., Waters, Robert Jordan, Ray, Jayashree, Carlson, Hans K., Lamson, Jacob S., Chakraborty, Romy, Arkin, Adam P., and Deutschbauer, Adam M. Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria. United States: N. p., 2018.
Web. doi:10.1128/mSystems.00143-17.
Liu, Hualan, Price, Morgan N., Waters, Robert Jordan, Ray, Jayashree, Carlson, Hans K., Lamson, Jacob S., Chakraborty, Romy, Arkin, Adam P., & Deutschbauer, Adam M. Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria. United States. https://doi.org/10.1128/mSystems.00143-17
Liu, Hualan, Price, Morgan N., Waters, Robert Jordan, Ray, Jayashree, Carlson, Hans K., Lamson, Jacob S., Chakraborty, Romy, Arkin, Adam P., and Deutschbauer, Adam M. Tue .
"Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria". United States. https://doi.org/10.1128/mSystems.00143-17. https://www.osti.gov/servlets/purl/1434009.
@article{osti_1434009,
title = {Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria},
author = {Liu, Hualan and Price, Morgan N. and Waters, Robert Jordan and Ray, Jayashree and Carlson, Hans K. and Lamson, Jacob S. and Chakraborty, Romy and Arkin, Adam P. and Deutschbauer, Adam M.},
abstractNote = {Molecular genetics is indispensable for interrogating the physiology of bacteria. However, the development of a functional genetic system for any given bacterium can be time-consuming. Here, we present a streamlined approach for identifying an effective transposon mutagenesis system for a new bacterium. Our strategy first involves the construction of hundreds of different transposon vector variants, which we term a “magic pool.” The efficacy of each vector in a magic pool is monitored in parallel using a unique DNA barcode that is introduced into each vector design. Using archived DNA “parts,” we next reassemble an effective vector for making a whole-genome transposon mutant library that is suitable for large-scale interrogation of gene function using competitive growth assays. Here, we demonstrate the utility of the magic pool system to make mutant libraries in five genera of bacteria.},
doi = {10.1128/mSystems.00143-17},
journal = {mSystems},
number = 1,
volume = 3,
place = {United States},
year = {Tue Jan 16 00:00:00 EST 2018},
month = {Tue Jan 16 00:00:00 EST 2018}
}
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