Document Detail


Optimization of electrotransformation conditions for Leuconostoc mesenteroides subsp. mesenteroides ATCC8293.
MedLine Citation:
PMID:  22897881     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Aims:  To establish an efficient genetic transformation protocol for Leuconostoc species, methods for competent-cell preparation and electroporation conditions were optimized. Methods and Results:  L. mesenteroides subsp. mesenteroides ATCC8293 cells were sequentially treated with penicillin G and lysozyme, and the plasmid pLeuCM was subsequently transformed into the cells. Our results demonstrated that transformation efficiencies were significantly increased (100-fold), and increased electric field strength also contributed to enhance transformation efficiency. Maximum transformation efficiency (1 × 10(4) or more transformants per μg DNA) was achieved when cells were grown in MRS media containing 0.25 M sucrose and 0.8 μg ml(-1) penicillin G, followed by treatment with 600 U ml(-1) lysozyme and electroporation at a field strength of 10 kV cm(-1) . When this protocol was used to transform pLeuCM into L. mesenteroides, L. gelidum, L. fallax, and L. argentinun, successful transformantions were obtained in all cases. Furthermore, this procedure was applicable to species belonging to other genera, including Lactobacillus plantarum, Pediococcus pentosaceus and Weissella confusa. Conclusions:  The results demonstrate that the transformation efficiency for Leuconostoc spp. could be increased via optimization of the entire electroporation procedures. © 2012 The Authors Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.
Authors:
Qing Jin; Hyun-Ju Eom; Jee Yun Jung; Jin Seok Moon; Jeong Hwan Kim; Nam Soo Han
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-8-16
Journal Detail:
Title:  Letters in applied microbiology     Volume:  -     ISSN:  1472-765X     ISO Abbreviation:  Lett. Appl. Microbiol.     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-8-17     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8510094     Medline TA:  Lett Appl Microbiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2012 The Authors Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.
Affiliation:
Department of Food Science, Yanbian University, Yanji, Jilin, China Department of Food Science and Technology, BK21 Education and Research Center for Advanced Bio-Agriculture Technology, Chungbuk National University, Cheongju, Korea Division of Applied Life Science (BK21), Graduate school, Gyeongsang National University, Jinju, Korea.
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