Summary
A plasmid, carrying the Tn5 gene for kanamycin resistance lacking its own promoter, has successfully been used in the selection of DNA sequences of the fungus Phycomyces blakesleeanus having promoter activity in Escherichia coli. Many of these sequences were also effective in promoting resistance to kanamycin when the corresponding chimeric plasmids were introduced in the fungus via spheroplast transformation. The selected phenotype was easily propagated through vegetative spores and behaved as a stable character since it was not appreciably lost in the absence of selection.
This is a preview of subscription content, log in via an institution to check access.
References
Bellofatto V, Shapiro L, Hodgson D (1984) Generation of a Tn5 promoter probe and its use in the study of gene expression in Caulobacter crescentus. Proc Natl Acad Sci USA 81:1035–1039
Bergman K, Eslava AP, Cerdá-Olmedo E (1973) Mutants of Phycomyces with abnormal phototropism. Mol Gen Genet 123:1–16
Buxton FP, Radford A (1984) The transformation of mycelial spheroplasts of Neurospora crassa and the attempted isolation of an autonomous replicator. Mol Gen Genet 196:339–344
Cerdá-Olmedo E, Lipson ED (1987) Phycomyces. Cold Spring Harbor Laboratory Press, New York
Eslava AP, Alvarez MI, Lipson ED, Presti D, Kong K (1976) Recombination between mutants of Phycomyces with abnormal phototropism. Mol Gen Genet 147:235–241
Hamer JE, Timberlake WE (1987) Functional organization of the Aspergillus nidulans trpC promoter. Mol Cell biol 7:2352–2359
López-Díaz I, Cerdá-Olmedo E (1980) Relationship of photocarotenogenesis to other behavioral and regulatory responses in Phycomyces. Planta 150:134–139
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Micol JL, Murillo FJ (1986) Characterization of Phycomyces blakesleeanus mutants temperature-sensitive for heat-shock induced germination. Curr Genet 10:755–760
Mishra NC (1985) Gene transfer in fungi. In: Caspari EW, Scandalios JG (eds) Advances in genetics, vol 23. Academic Press, New York, p 74–178
Murillo FJ, Calderón IL, López-Díaz I, Cerdá-Olmedo E (1978) Carotene-superproducing strains of Phycomyces. Appl Environ Microbiol 36:639–642
Neve RL, West RW, Rodríguez RL (1979) Eukaryotic DNA fragments which act as promoters for a plasmid gene. Nature 277:324–325
Revuelta JL, Jayaram M (1986) Transformation of Phycomyces blakesleeanus to G-418 resistance by an autonomously replicating plasmid. Proc Natl Acad Sci USA 83:7344–7347
Revuelta JL, Jayaram M (1987) Phycomyces blakesleeanus TRP1 gene: Organization and functional complementation in Escherichia coli and Saccharomyces cerevisiae. Mol Cell Biol 7:2664–2670
Roncero MIG, Zabala C, Cerdá-olmedo E (1984) Mutagenesis in multinucleate cells: The effects of N-methyl-N′-nitro-N-nitrosoguanidine in Phycomyces spores. Mutat Res 125:195–204
Suárez T (1985) Obtención de protoplastos y transformación en Phycomyces blakesleeanus. Tesis Doctoral, Universidad de Salamanca, España
Suárez T, Orejas M, Arnau J, Torres-Martínez S (1987) Protoplast formation and fusion. In: Cerdá-Olmedo E, Lipson ED (eds) Phycomyces. Cold Spring Harbor Laboratory Press, New York, pp 351–353
Sutter RP (1975) Mutations affecting sexual development in Phycomyces blakesleeanus. Proc Natl Acad Sci USA 72:127–130
Van Heeswijck R, Roncero MIG (1984) High frequency transformation of Mucor with recombinant plasmid DNA. Carlsberg Res Commun 49:691–702
Author information
Authors and Affiliations
Additional information
Communicated by W. Gajewski
Rights and permissions
About this article
Cite this article
Arnau, J., Murillo, F.J. & Torres-Martínez, S. Expression of Tn5-derived kanamycin resistance in the fungus Phycomyces blakesleeanus . Mol Gen Genet 212, 375–377 (1988). https://doi.org/10.1007/BF00334710
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00334710