| 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 |
Related Documents
:
|
10049249 - Reca-mediated gene conversion and aminoglycoside resistance in strains heterozygous for... 12501389 - Transformation with green fluorescent protein of trichoderma harzianum 1051, a strain w... 1905659 - Genetic transformation of various species of enterococcus by electroporation. 16410769 - Screening of growth- or development-related genes by using genomic library with inducib... 137229 - Mutations in prophage phi11 that impair the transducibility of their staphylococcus aur... 2988449 - Cloning and expression of the mete gene in escherichia coli. 11478669 - Sensitivity and performance characteristics of a direct pcr with stool samples in compa... 20081009 - Hwanghaeicola aestuarii gen. nov., sp. nov., a moderately halophilic bacterium isolated... 9557319 - Comparison of the protein patterns of erysipelothrix rhusiopathiae strains by sds-page ... |
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. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| 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
Previous Document: Development of surgical skill with singular neurectomy using human cadaveric temporal bones.
Next Document: Synergism between natural products and antibiotics against infectious diseases.