Document Detail


An efficient genetic transformation method for glycerol producer Candida glycerinogenes.
MedLine Citation:
PMID:  18599279     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Transformation techniques generally require development before genetic and molecular studies of industrial yeast strains can commence. Candida glycerinogenes WL2002-5 has been used for industrial-scale glycerol production but has proven difficult to transform for molecular studies following previously published procedures. In the present study, phleomycin was used as drug-resistance marker based on perturbing plasma membranes and the lower minimum inhibitory concentration for C. glycerinogenes. We developed an efficient method for transformation of C. glycerinogenes, and parameters involved in transformation efficiency were optimized. Pretreatment of yeast cells with either dithiothreitol or lithium acetate enhanced the frequency of transformation markedly by electroporation. Significantly higher transformation efficiency was obtained when the electroporated cells were pretreated with phleomycin before plating. With this method, a maximal transformation frequency of 394+/-50transformants/mug plasmid DNA was obtained. The transformation method will foster genetic manipulation of this industrial strain.
Authors:
Xianzhong Chen; Huiying Fang; Zhiming Rao; Wei Shen; Bin Zhuge; Zhengxiang Wang; Jian Zhuge
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Publication Detail:
Type:  Evaluation Studies; Journal Article     Date:  2008-07-02
Journal Detail:
Title:  Microbiological research     Volume:  163     ISSN:  0944-5013     ISO Abbreviation:  Microbiol. Res.     Publication Date:  2008  
Date Detail:
Created Date:  2008-08-12     Completed Date:  2008-10-09     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9437794     Medline TA:  Microbiol Res     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  531-7     Citation Subset:  IM    
Affiliation:
The Key Lab of Industrial Biotechnology, The Ministry of Education and Research Center of Industrial Microorganisms, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
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MeSH Terms
Descriptor/Qualifier:
Acetates / pharmacology
Anti-Bacterial Agents / pharmacology
Candida / drug effects,  genetics*,  metabolism
Dithiothreitol / pharmacology
Gene Transfer Techniques*
Genome, Fungal
Glycerol / metabolism*
Industrial Microbiology*
Plasmids / genetics
Transformation, Genetic*
Chemical
Reg. No./Substance:
0/Acetates; 0/Anti-Bacterial Agents; 0/lithium acetate; 3483-12-3/Dithiothreitol; 56-81-5/Glycerol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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