Construction of an autonomously replicating plasmid in n-alkane- assimilating yeast, Candida tropicalis

Akihiro Hara, Mitsuyoshi Ueda, Toru Matsui, Keizo Furuhashi, Naoki Kanayama, Atsuo Tanaka

Research output: Contribution to journalArticle

7 Citations (Scopus)

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 languageEnglish
Pages (from-to)717-720
Number of pages4
JournalJournal of Bioscience and Bioengineering
Volume87
Issue number6
DOIs
Publication statusPublished - Jan 1 1999
Externally publishedYes

Keywords

  • Autonomously replicating sequence (ARS)
  • Candida tropicalis
  • Dicarboxylic acid
  • n- alkane

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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