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Cloning and characterization of a gene encoding a malic enzyme involved in anaerobic growth in Mucor circinelloides

Published online by Cambridge University Press:  30 March 2005

Yonghua LI
Affiliation:
Centre for Lipid Research, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. E-mail: [email protected] Present address: Department of Plant Biology, Michigan State University, East Lansing, MI 48823, USA.
Ian P. ADAMS
Affiliation:
Centre for Lipid Research, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. E-mail: [email protected] Present address: Division of Academic Medicine, Postgraduate Medical School, University of Hull, Cottingham Road, Hull HU6 7RX, UK.
James P. WYNN
Affiliation:
Centre for Lipid Research, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. E-mail: [email protected] Present address: Martek Biosciences Corporation, 6480 Dobbin Road, Columbia, MD 21045, USA.
Colin RATLEDGE
Affiliation:
Centre for Lipid Research, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. E-mail: [email protected]
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Abstract

A 3193 bp contiguous sequence has been cloned from the oleaginous fungus Mucor circinelloides, that contains a full-length gene encoding a putative NADP+[ratio ]dependent malic enzyme (EC. 1.1.1.40). The cloned DNA contains a 2154 bp putative open reading frame containing five introns and encoding a protein of 616 amino acids. The gene encoded what appeared to be an anaerobic isoform of malic enzyme (isoform II); this conclusion is supported by transcript analysis and by the fact that the ORF contains an N-terminal mitochondrial target sequence (a similar cellular location was identified for the anaerobic malic enzyme in Saccharomyces cerevisiae; Boles et al. 1998). The cloned gene did not encode either isoform III (the isoform associated with active growth) or isoform IV (associated with lipid accumulation) previously identified. M. circinelloides therefore must possess (at least) two structural genes for malic enzyme.

Type
Research Article
Copyright
© The British Mycological Society 2005

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