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Appl Environ Microbiol, July 1998, p. 2624-2629, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cloning of Phanerochaete chrysosporium leu2 by Complementation of Bacterial Auxotrophs and Transformation of Fungal Auxotrophs

Laura Schick Zapanta, Takefumi Hattori,dagger Magarita Rzetskaya,Dagger and Ming Tien*

Department of Biochemistry and Molecular Biology and The Center for Biomolecular Structure and Function, The Pennsylvania State University, University Park, Pennsylvania 16802

Received 25 September 1997/Accepted 27 February 1998

A Phanerochaete chrysosporium cDNA library was constructed in an expression vector that allows expression in both Escherichia coli and Saccharomyces cerevisiae. This expression vector, lambda YES, contains the lacZ promoter for expression in E. coli and the GAL1 promoter for expression in yeast. A number of genes were cloned by complementation of bacterial amino acid auxotrophs. The cDNA encoding the beta -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.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, 408 Althouse Laboratory, The Pennsylvania State University, University Park, PA 16802. Phone: (814) 863-1165. Fax: (814) 863-8616. E-mail: mxt3{at}psu.edu.

dagger Present address: Wood Research Institute, Kyoto University, Kyoto, Japan.

Dagger Present address: Columbia University, New York, NY 10032.

Appl Environ Microbiol, July 1998, p. 2624-2629, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.




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