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PMID: 8501059 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

High-frequency transformation of a methylotrophic yeast, Candida boidinii, with autonomously replicating plasmids which are also functional in Saccharomyces cerevisiae.

Journal of bacteriology ·Vol. 175 ·No. 11 ·1993-06-00 ·Pages 3556-62

Sakai Y, Goh TK, Tani Y

Abstract

We have developed a transformation system which uses autonomous replicating plasmids for a methylotrophic yeast, Candida boidinii. Two autonomous replication sequences, CARS1 and CARS2, were newly cloned from the genome of C. boidinii. Plasmids having both a CARS fragment and the C. boidinii URA3 gene transformed C. boidinii ura3 cells to Ura+ phenotype at frequencies of up to 10(4) CFU/micrograms of DNA. From Southern blot analysis, CARS plasmids seemed to exist in polymeric forms as well as in monomeric forms in C. boidinii cells. The C. boidinii URA3 gene was overexpressed in C. boidinii on these CARS vectors. CARS1 and CARS2 were found to function as an autonomous replicating element in Saccharomyces cerevisiae as well. Different portions of the CARS1 sequence were needed for autonomous replicating activity in C. boidinii and S. cerevisiae. C. boidinii could also be transformed with vectors harboring a CARS fragment and the S. cerevisiae URA3 gene.

Related Genes
URA3
MeSH Terms
Acetates/pharmacology Acetic Acid Base Sequence Candida/genetics Cloning, Molecular DNA Replication Escherichia coli/genetics Genes, Fungal/genetics Genetic Vectors/genetics Methanol/metabolism Molecular Sequence Data Orotidine-5'-Phosphate Decarboxylase/biosynthesis,genetics Phenotype Plasmids/genetics Saccharomyces cerevisiae/genetics Transformation, Genetic/drug effects
Chemicals
Acetates Orotidine-5'-Phosphate Decarboxylase Acetic Acid Methanol
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sakai Y
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.
Goh T K
Tani Y
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39 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.175.11.3556-3562.1993
Published
1993-06-00
Pages
3556-62
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC204756
Subset
IM
Databases
GENBANK
L13785
External Links
PubMed source DOI: 10.1128/jb.175.11.3556-3562.1993 PMC Full Text View journal details
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