Abstract
A transformation system using the autonomously replicating plasmid in the n-alkane-assimilating and asporogenic diploid yeast, Candida tropicalis, was developed. For the cloning of a DNA fragment containing a potential autonomously replicating sequence (ARS) from the genomic DNA of C. tropicalis, the ura3 mutant obtained using ethylmethane sulfonate as the host and the URA3 gene amplified by PCR using the C. tropicalis genomic DNA as a selectable marker were prepared. Comparison of ARSs among yeasts revealed that the consensus sequence found in S. cerevisiae was also present in C. tropicalis. The autonomously replicating plasmid containing the putative ARS as the shuttle vector, capable of replicating in both E. coli and C. tropicalis, was first constructed. The transformation system using this plasmid, in addition to the integrative transformation system, will be applicable to genetic studies of C. tropicalis.
| Original language | English |
|---|---|
| Pages (from-to) | 717-720 |
| Number of pages | 4 |
| Journal | Journal of Bioscience and Bioengineering |
| Volume | 87 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 1999 |
| Externally published | Yes |
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Keywords
- Autonomously replicating sequence (ARS)
- Candida tropicalis
- Dicarboxylic acid
- n- alkane
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
Cite this
Construction of an autonomously replicating plasmid in n-alkane- assimilating yeast, Candida tropicalis. / Hara, Akihiro; Ueda, Mitsuyoshi; Matsui, Toru; Furuhashi, Keizo; Kanayama, Naoki; Tanaka, Atsuo.
In: Journal of Bioscience and Bioengineering, Vol. 87, No. 6, 1999, p. 717-720.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Construction of an autonomously replicating plasmid in n-alkane- assimilating yeast, Candida tropicalis
AU - Hara, Akihiro
AU - Ueda, Mitsuyoshi
AU - Matsui, Toru
AU - Furuhashi, Keizo
AU - Kanayama, Naoki
AU - Tanaka, Atsuo
PY - 1999
Y1 - 1999
N2 - A transformation system using the autonomously replicating plasmid in the n-alkane-assimilating and asporogenic diploid yeast, Candida tropicalis, was developed. For the cloning of a DNA fragment containing a potential autonomously replicating sequence (ARS) from the genomic DNA of C. tropicalis, the ura3 mutant obtained using ethylmethane sulfonate as the host and the URA3 gene amplified by PCR using the C. tropicalis genomic DNA as a selectable marker were prepared. Comparison of ARSs among yeasts revealed that the consensus sequence found in S. cerevisiae was also present in C. tropicalis. The autonomously replicating plasmid containing the putative ARS as the shuttle vector, capable of replicating in both E. coli and C. tropicalis, was first constructed. The transformation system using this plasmid, in addition to the integrative transformation system, will be applicable to genetic studies of C. tropicalis.
AB - A transformation system using the autonomously replicating plasmid in the n-alkane-assimilating and asporogenic diploid yeast, Candida tropicalis, was developed. For the cloning of a DNA fragment containing a potential autonomously replicating sequence (ARS) from the genomic DNA of C. tropicalis, the ura3 mutant obtained using ethylmethane sulfonate as the host and the URA3 gene amplified by PCR using the C. tropicalis genomic DNA as a selectable marker were prepared. Comparison of ARSs among yeasts revealed that the consensus sequence found in S. cerevisiae was also present in C. tropicalis. The autonomously replicating plasmid containing the putative ARS as the shuttle vector, capable of replicating in both E. coli and C. tropicalis, was first constructed. The transformation system using this plasmid, in addition to the integrative transformation system, will be applicable to genetic studies of C. tropicalis.
KW - Autonomously replicating sequence (ARS)
KW - Candida tropicalis
KW - Dicarboxylic acid
KW - n- alkane
UR - http://www.scopus.com/inward/record.url?scp=0032778702&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032778702&partnerID=8YFLogxK
U2 - 10.1016/S1389-1723(99)80143-1
DO - 10.1016/S1389-1723(99)80143-1
M3 - Article
C2 - 16232544
AN - SCOPUS:0032778702
VL - 87
SP - 717
EP - 720
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
SN - 1389-1723
IS - 6
ER -