Genetic Introduction of Foreign Genes to Pleurotus eryngii by Restriction Enzyme-Mediated Integration

Won Noh; Sang-Woo Kim; Dong-Won Bae; Jae-Yean Kim; Hyeon-Su Ro

doi:10.1007/s12275-010-9278-7

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Research Support, Non-U.S. Gov't
Genetic Introduction of Foreign Genes to Pleurotus eryngii by Restriction Enzyme-Mediated Integration: Won Noh ¹, Sang-Woo Kim ¹, Dong-Won Bae ², Jae-Yean Kim ³, Hyeon-Su Ro ^1,3; Journal of Microbiology 2010;48(2):253-256
DOI: https://doi.org/10.1007/s12275-010-9278-7
Published online: May 1, 2010

¹Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, Chinju 660-701, Republic of Korea, ²Central Instrument Facility, Gyeongsang National University, Chinju 660-701, Republic of Korea, ³Division of Applied Life Science and Environmental Biotechnology National Core Research Center, Gyeongsang National University, Chinju 660-701, Republic of Korea¹Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, Chinju 660-701, Republic of Korea, ²Central Instrument Facility, Gyeongsang National University, Chinju 660-701, Republic of Korea, ³Division of Applied Life Science and Environmental Biotechnology National Core Research Center, Gyeongsang National University, Chinju 660-701, Republic of Korea

Corresponding author: Hyeon-Su Ro , Tel: +82-55-751-5946,

Received: 13 September 2009 • Accepted: 15 October 2009

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Abstract

Pleurotus eryngii was transformed via restriction enzyme-mediated integration. In order to construct the transformation plasmid, the enhanced cyan fluorescent protein (ECFP) gene was ligated next to the gpd promoter of the plasmid pAN7-1. Transformation was facilitated via the heat treatment of a transformation mixture containing 1 μg of the HindIII-digested plasmid DNA and 106 mushroom protoplasts in 40% polyethyleneglycol solution, resulting in 10-40 hygromycin-resistant transformants. Successful transformation was evidenced by PCR, Southern blot, and confocal fluorescence microscopic analyses on the selected transformants. To date, this is the first report on the transformation of P. eryngii by REMI technique.

Keywords: hygromycin; P. eryngii; REMI; transformation

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Genetic Introduction of Foreign Genes to Pleurotus eryngii by Restriction Enzyme-Mediated Integration

J. Microbiol. 2010;48(2):253-256. Published online May 1, 2010

DOI: https://doi.org/10.1007/s12275-010-9278-7

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