Cloning of Phanerochaete chrysosporium leu2 by complementation of bacterial auxotrophs and transformation of fungal auxotrophs

Laura Schick Zapanta, Takefumi Hattori, Magarita Rzetskaya, Ming Tien

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

A Phanerochaete chrysosporium cDNA library was constructed in an expression vector that allows expression in both Escherichia coli and Saccharomyces cerevisiae. This expression vector, λYES, contains the lacZ promoter for expression in E. coli and the GALl promoter for expression in yeast. A number of genes were cloned by complementation of bacterial amino acid auxotrophs. The cDNA encoding the β-isopropylmalate dehydrogenase from P. chrysosporium was characterized further. The genomic clone (gleu2) was subsequently isolated and was used successfully as a selectable marker to transform P. chrysosporium auxotrophs for LEU2. Protoplasts for transformation were prepared with readily obtained conidiospores rather than with basidiospores, which were used in previous P. chrysosporium transformation procedures. The method described here allows other genes to be isolated from P. chrysosporium for use as selectable markers.

Original languageEnglish (US)
Pages (from-to)2624-2629
Number of pages6
JournalApplied and Environmental Microbiology
Volume64
Issue number7
StatePublished - Jul 1998

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Bacterial Transformation
Phanerochaete
Phanerochaete chrysosporium
auxotrophs
Organism Cloning
molecular cloning
gene
yeast
clone
genomics
transform
amino acid
promoter regions
Escherichia coli
basidiospores
Protoplasts
Gene Library
cDNA libraries
protoplasts
Genes

All Science Journal Classification (ASJC) codes

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

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Cloning of Phanerochaete chrysosporium leu2 by complementation of bacterial auxotrophs and transformation of fungal auxotrophs. / Zapanta, Laura Schick; Hattori, Takefumi; Rzetskaya, Magarita; Tien, Ming.

In: Applied and Environmental Microbiology, Vol. 64, No. 7, 07.1998, p. 2624-2629.

Research output: Contribution to journalArticle

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AU - Tien, Ming

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