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Bioscience, Biotechnology, and Biochemistry Volume 76, 2012 - Issue 2
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Original Articles

Isolation of Uracil Auxotrophs of the Fungus Acremonium cellulolyticus and the Development of a Transformation System with the pyrF Gene

Tatsuya FUJIIBiomass Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST)
,
Kazuya IWATABiomass Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST)
,
Katsuji MURAKAMIBiomass Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST)
,
Shinichi YANOBiomass Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST)
&
Shigeki SAWAYAMABiomass Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST);Division of Applied Bioscience, Graduate School of Agriculture, Kyoto University
Pages 245-249 | Received 04 Jul 2011, Accepted 27 Oct 2011, Published online: 22 May 2014
 
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Abstract

Acremonium cellulolyticus CF-2612 is a cellulase hyper-producing mutant that originated from A. cellulolyticus Y-94. In this study, we isolated a uracil auxotroph (strain CFP3) derived from CF-2612, and cloned a wild-type pyrF gene encoding orotate phosphoribosyl transferase (OPRTase) from Y-94. OPRTase activity was not detected in strain CFP3, which had one nucleotide substitution in its pyrF gene. The wild-type pyrF gene restored the defective growth of CFP3 on uracil-free medium, and PCR and Southern analyses revealed that wild-type pyrF was integrated into the genome. These results indicate that our transformation system for A. cellulolyticus with the pyrFgene as a selection marker was successful.

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Related Research Data

Structural and functional characterization of a bifunctional GH30-7 xylanase B from the filamentous fungus Talaromyces cellulolyticus
Source: American Society for Biochemistry & Molecular Biology (ASBMB)
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The Putative Transcription Factor Gene thaB Regulates Cellulase and Xylanase Production at the Enzymatic and Transcriptional Level in the Fungus Talaromyces cellulolyticus.
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