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Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria
Korean J. Food Sci. An. 2018;38:816-822
Published online August 31, 2018;  https://doi.org/10.5851/kosfa.2018.e17
© 2018 Korean Society for Food Science of Animal Resources

Gi-Seong Moon1,* and Arjan Narbad2

1Department of Biotechnology, Korea National University of Transportation, Jeungpyeong 27909, Korea
2Translational Microbiome (Narbad Group), Quadram Institute Bioscience, Norwich NR4 7UA, UK
Correspondence to: Gi-Seong Moon
Department of Biotechnology, Korea National University of Transportation, Jeungpyeong 27909, Korea
Tel: +82-43-820-5251
Fax: +82-43-820-5272
E-mail: gsmoon@ut.ac.kr
Received June 11, 2018; Revised July 6, 2018; Accepted July 6, 2018.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Bifidobacterium is recognized as one of the most beneficial microorganisms in our gut. Many researches on bifidobacteria have been done to understand their roles in the gut. The objective of the present study was to develop a bioluminescent labelling plasmid vector for bifidobacteria to facilitate their visualization in vitro, in situ, and in vivo. A plasmid replicon (2.0 kb) of plasmid pFI2576 previously identified from B. longum FI10564 was amplified by PCR and cloned into pUC19 plasmid vector (2.68 kb). The cloned replicon was subcloned into pTG262 (luc+) recombinant plasmid vector (7.4 kb) where a luciferase gene (luc+) from pLuc2 (8.5 kb), an Escherichia coli and lactobacilli shuttle vector, was inserted into pTG262 plasmid vector. The final recombinant DNA, pTG262::pFI2576 rep (luc+), was transferred into a B. catenulatum strain. This recombinant strain showed 3,024 relative luminescence units at OD600 value of 0.352. Thus, this recombinant plasmid construct can be broadly used for labelling bifidobacteria.
Keywords : Bifidobacterium, bioluminescence, luciferase gene, plasmid vector, replicon


October 2018, 38 (5)