Novel Bifidobacterium Promoters Selected Through Microarray Analysis Lead to Constitutive High-Level Gene Expression

Yan Wang; Jin Yong Kim; Myeong Soo Park; Geun Eog Ji

doi:10.1007/s12275-012-1591-x

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Novel Bifidobacterium Promoters Selected Through Microarray Analysis Lead to Constitutive High-Level Gene Expression: Yan Wang ¹, Jin Yong Kim ¹, Myeong Soo Park ^2,3, Geun Eog Ji ^1,3; Journal of Microbiology 2012;50(4):638-643.
DOI: https://doi.org/10.1007/s12275-012-1591-x
Published online: July 21, 2012

¹Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul 151-742, Republic of Korea, ²Department of Hotel Culinary Arts, Anyang Science University, Anyang 430-749, Republic of Korea, ³Research Institute, BIFIDO Co., Ltd. Hongchun 250-804, Republic of Korea¹Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul 151-742, Republic of Korea, ²Department of Hotel Culinary Arts, Anyang Science University, Anyang 430-749, Republic of Korea, ³Research Institute, BIFIDO Co., Ltd. Hongchun 250-804, Republic of Korea

Corresponding author: Myeong Soo Park , Tel: +82-2-880-8749/+82-31-441-1347,
Geun Eog Ji , Tel: +82-2-880-8749/+82-31-441-1347,

Received: 7 December 2011 • Accepted: 17 April 2012

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Abstract

For the development of a food-grade expression system for Bifidobacterium, a strong promoter leading to high-level expression of cloned gene is a prerequisite. For this purpose, a promoter screening host-vector system for Bifidobacterium has been established using β-glucosidase from Bifidobacterium lactis as a reporter and Bifidobacterium bifidum BGN4 as a host, which is β-glucosidase negative strain. Seven putative promoters showing constitutive high-level expression were selected through microarray analysis based on the genome sequence of B. bifidum BGN4. They were cloned into upstream of β-glucosidase gene and transformed into Escherichia coli DH5α and B. bifidum BGN4. Promoter activities were analyzed both in E. coli and B. bifidum BGN4 by measuring β-glucosidase activity. β-Glucosidase activities in all of the transformants showed growth-associated characteristics. Among them, P919 was the strongest in B. bifidum BGN4 and showed maximum activity at 18 h, while P895 was the strongest in E. coli DH5α at 7 h. This study shows that novel strong promoters such as P919 can be used for high-level expression of foreign genes in Bifidobacterium and will be useful for the construction of an efficient food-grade expression system.

Keywords: Bifidobacterium;promoter;expression;vector

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Novel Bifidobacterium Promoters Selected Through Microarray Analysis Lead to Constitutive High-Level Gene Expression

J. Microbiol. 2012;50(4):638-643. Published online July 21, 2012

DOI: https://doi.org/10.1007/s12275-012-1591-x

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