Abstract
The sdhB gene encoding an iron–sulfur (Ip) subunit of succinate dehydrogenase (SDH, EC 1.3.99.1) complex was cloned from Mortierella alpina 1S-4. The deduced amino acid sequence of SdhB from M. alpina 1S-4 showed high similarity to those of SdhB from other organisms. The mutated sdhB (CBXB) gene encodes a modified SdhB with an amino-acid substitution (a highly conserved histidine residue within the third cysteine-rich cluster of SdhB replaced by a leucine residue) and is known to confer carboxin resistance. We succeeded in transforming M. alpina 1S-4 by using the CBXB gene as a selectable marker gene and expressing the heterologous uidA gene encoding β-glucuronidase of Escherichia coli. Moreover, transformation efficiency was up to 40–50 transformants per 4.0 × 108 spores. This carboxin-transformation system, characterized by marginal background growth and mitotic stability in M. alpina 1S-4, is considered to be widely useful for the wild strain, M. alpina 1S-4, and various derivative mutants without laborious preparation of auxotrophic mutants as a host strain.
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Acknowledgments
This work was partially supported by the Project for the Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers (to S.S.), the Industrial Technology Research Grant Program (No. 05A07003d to E.S.) of the New Energy and Industrial Technology Development Organization, Japan, Grants-in-Aid for Scientific Research (No. 16688004 to J.O. and No. 19688006 to E.S.), and COE for Microbial-Process Development Pioneering Future Production Systems (to S.S.) from the Ministry of Education, Science, Sports, and Culture, Japan.
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Communicated by U. Kueck.
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Ando, A., Sakuradani, E., Horinaka, K. et al. Transformation of an oleaginous zygomycete Mortierella alpina 1S-4 with the carboxin resistance gene conferred by mutation of the iron–sulfur subunit of succinate dehydrogenase. Curr Genet 55, 349–356 (2009). https://doi.org/10.1007/s00294-009-0250-1
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DOI: https://doi.org/10.1007/s00294-009-0250-1