Summary
An Aspergillus nidulans gene library was constructed in a high-frequency transformation vector, pDJB3, based on the Neurospora crassa pyr4 gene. This gene library was used to isolate the structural gene for isocitrate lyase (acuD) by complementation of a deficiency mutation following transformation of A. nidulans. Plasmids rescued in Escherichia coli were able to transform five different A. nidulans acuD mutants. Transformation using plasmids containing the cloned fragment resulted in integration at the acuD locus in six of nine transformants.
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References
Apirion D (1965) The two-way selection of mutants and revertants in respect of acetate utilisation and resistance to fluoroacetate in Aspergillus nidulans. Genet Res 6:317–329
Armitt S, McCullough W, Roberts CF (1976) Analysis of acetate non-utilizing (acu) mutants in Aspergillus nidulans. J Gen Microbiol 92:263–282
Ballance DJ, Turner G (1985) Development of a high-frequency transforming vector for Aspergillus nidulans. Gene 36:321–331
Ballance DJ, Buxton FP, Turner G (1983) Transformation of Aspergillus nidulans by the orotidine-5′-phosphate decarboxylase gene of Neurospora crassa. Biochem Biophys Res Commun 112:284–289
Dretzen G, Bellard M, Sassone-Corsi P, Chambon P (1981) A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem 112:295–298
Edwards GFStL, Holt G, Macdonald KD (1974) Mutants of Aspergillus nidulans impaired in penicillin biosynthesis. J Gen Microbiol 84:420–422
Hanahan D (1982) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci USA 75:1929–1933
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Hynes MJ (1977) Induction of the acetamidase of Aspergillus nidulans by acetate metabolism. J Bacteriol 131:770–775
Hynes MJ, Corrick CM, King JA (1983) Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol Cell Biol 3:1430–1439
Johanson RA, Hill JM, McFadden BA (1974) Isocitrate lyase from Neurospora crassa II. Composition, quanternary structure, C-terminus, and active-site modification. Biochim Biophys Acta 364:341–352
Johnstone JL, Hughes SG, Clutterbuck AJ (1985) Cloning of an Aspergillus nidulans developmental gene by transformation. EMBO J 4:1307–1311
Kelly JM, Hynes MJ (1985) Transformation of Aspergillus niger by the amdS gene of Aspergillus nidulans. EMBO J 4:475–479
Kornberg HL (1966) The role and control of the glyoxylate cycle in Escherichia coli. Biochem J 99:1–11
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
Marinus MG, Konrad EB (1976) Hyper-recombination in dam mutants of Escherichia coli K-12. Mol Gen Genet 149:273–277
Palmer LM, Cove DJ (1975) Pyrimidine biosynthesis in Aspergillus nidulans. Isolation and preliminary characterisation of auxotrophic mutants. Mol Gen Genet 138:243–255
Rowlands RT, Turner G (1977) Nuclear and extranuclear inheritance in Aspergillus nidulans. Mol Gen Genet 126:201–216
Rowlands RT, Turner G (1977) Nuclear-extranuclear interactions affecting oligomycin resistance in Aspergillus nidulans. Mol Gen Genet 154:311–318
Tilburn J, Scazzocchio C, Taylor GG, Zabicky-Zissman JH, Lockington RA, Davies RW (1983) Transformation by integration in Aspergillus nidulans. Gene 26:205–221
Twigg AJ, Sherratt D (1980) Trans-complementable copy-number mutants of plasmid ColE1. Nature 283:216–218
Weiss RL, Puetz D, Cybis J (1985) Expression of Aspergillus genes in Neurospora. In: Bennett JW, Lasure LL (eds) Gene manipulations in fungi. Academic Press, New York, in press
Yelton MM, Hamer JE, Timberlake WE (1984) Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci USA 81:1470–1474
Yelton MM, Timberlake WE, van den Hondel CAMJJ (1985) A cosmid for selecting genes by complementation in Aspergillus nidulans: Selection of the developmentally regulated yA locus. Proc Natl Acad Sci USA 82:834–838
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Communicated by C.P. Hollenberg
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Ballance, D.J., Turner, G. Gene cloning in Aspergillus nidulans: isolation of the isocitrate lyase gene (acuD). Mol Gen Genet 202, 271–275 (1986). https://doi.org/10.1007/BF00331649
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DOI: https://doi.org/10.1007/BF00331649