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Received February 9, 2017
Accepted March 16, 2017
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This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits
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Development of Cre-lox based multiple knockout system in Deinococcus radiodurans R1
School of Environmental Engineering, University of Seoul, Seoul 02504, Korea 1Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Korea 2Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Gwahak-ro 125, Yuseong-gu, Daejeon 34141, Korea
Korean Journal of Chemical Engineering, June 2017, 34(6), 1728-1733(6), 10.1007/s11814-017-0082-5
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Abstract
The extremophilic bacterium Deinococcus radiodurans R1 has been considered as an attractive microorganism due to its remarkable tolerance to various external stresses. Considering the nature of D. radiodurans R1, it has potential as a platform microorganism for industrial applications, including biorefinery and bioremediation process. However, D. radiodurans R1 is well known for its hard genetic manipulation. Thus, much effort has been made to develop efficient genetic engineering tools for making D. radiodurans R1 suitable for industrial platform microorganism. Although a plasmid-based single gene knockout method has been reported, development of multiple gene knockout system has not yet been reported. Here we report, for the first time, Cre-lox based rapid and efficient multiple knockout method for metabolic engineering of D. radiodurans R1. Also, deletion of dr0053 gene was successfully achieved within seven days to make biofilm overproducing strain.
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References
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Meima R, Lidstrom ME, Appl. Environ. Microbiol., 66, 3856 (2000)
Nguyen HH, de la Tour CB, Toueille M, Vannier F, Sommer S, Servant P, Mol. Microbiol., 73, 240 (2009)
Satoh K, Tu Z, Ohba H, Narumi I, Plasmid, 62, 1 (2009)
Appukuttan D, Seo HS, Jeong SW, Im SH, Joe MH, Song DS, Choi JJ, Lim SY, PLoS One, 10, e0118275 (2015).
O’Toole GA, Pratt LA, Watnick PI, Newman DK, Weaver VB, Kolter RG, Methods Enzymol., 310, 91 (1999)
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Moseley BE, Setlow JK, Proc. Natl. Acad. Sci. U.S.A., 61, 176 (1968)
Lambert JM, Bonger RS, Kleerebezem M, Appl. Environ. Microbiol., 73, 11262 (2007)
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Kolari M, Nuutinen J, Salkinoja-Salonen MS, J. Ind. Microbiol. Biotechnol., 27, 343 (2001)
Slade D, Radman M, Microbiol. Mol. Biol. Rev., 75, 133 (2011)
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