Summary
In order to establish a transformation system for P. chrysogenum autonomously replicating vectors were constructed using mitochondrial DNA sequences from the fungus. A physical map of the mt DNA of a production strain was established using ten different restriction enzymes. Unexpectedly, the mt DNA of this strain proved to be significantly smaller than that of a second strain from a culture collection (27 kb versus 49 kb). Various fragments representing about 71% of the 27 kb mt DNA were cloned and, at first, preselected for replicating activity in an intermediate host (Saccharomyces cerevisiae). Two of these fragments also promoted autonomous replication in P. chrysogenum, which was confirmed by isolation of bulk DNA and transfer into E. coli. For selection of transformants in P. chrysogenum the prokaryotic kanamycin resistance gene was used which increased about twofold the resistance against G418.
Similar content being viewed by others
References
Ball C (1985) Perspectives for protoplasts in the genetics of industrially important fungi. In Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Mucology Series, Vol. 6, Marcel Dekker, New York-Basel, pp 337–343
Beach D, Piper M, Shall S (1980) Isolation of chromosomal origins of replication in yeast. Nature 284:185–187
Bolivar F (1978) Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique EcoR I sites for selection of EcoR I generated recombinant DNA molecules. Gene 4:121–136
Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P 15 A cryptic miniplasmid. J Bacteriol 134:1141–1156
Crueger WA, Crueger A (1982) Lehrbuch der angewandten Mikrobiologie. Akademische Verlagsgesellschaft, Wiesbaden
Esser K, Kück U, Stahl U, Tudzynski P (1983) Cloning vectors of mitochondrial origin for eukaryotes: a new concept in genetic engineering. Curr Genet 7:239–243
Esser K, Kück P, Lang-Hinrichs C, Lemke P, Osiewacz DH, Stahl U, Tudzynski P (1986) Plasmids of eukaryotes. Springer, Berlin Heidelberg New York Tokyo
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci USA 75:1929–1933
Hollenberg CP (1979) The expression of bacterial resistance genes in the yeast Saccharomyces cerevisiae. In: Timmis KN Pühler A (eds) Plasmids of medical, environmental and commercial importance. pp 481–492. Elsevier/North Holland Biomedical Press, Amsterdam-New York-Oxford
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Kück U, Kappelhoff B, Esser K (1985) Despite mt DNA polymorphism the mobile intron (pl DNA) of the COI gene is present in ten different races of Podospora anserina. Curr Genet 10:59–67
Lang-Hinrichs C (1985) Extrachromosomale in-vitro-Genetik bei Pilzen; Chondriom-Vektoren bei Hefen. Dissertation Bochum 1985
Lang-Hinrichs C, Stahl U (1987) Cloning vectors of mitochondrial origin for eukaryotes. In: Wu R (ed) Methods in enzymology. Academic Press, New York London (in press)
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning; Laboratory manual. Cold Spring Harbour
Rechcigl M (1978) CRC Handbook series in nutrition and food, Section G, Vol. III. CRC Press, Cleveland
Smith TM, Saunders G, Stacey LM, Holt G (1984) Restriction endonuclease map of mitochondrial DNA from Penicillium chrysogenum. J Biotechnol 1:37–46
Stahl U, Kück U, Tudzynski P, Esser K (1980) Characterization and cloning of plasmid like DNA of the ascomycete Podospora anserina. Mol Gen Genet 178:639–646
Stohl LL, Lambowitz AM (1983) Construction of a shuttle vector for the filamentous fungus Neuropora crassa. Proc Natl Acad Sci USA 80:1058–1062
Tilburn J, Scazzocchio C, Taylor GG, Zabicky-Zissman JH, Lockington RA, Davies RW (1983) Transformation by integration in Aspergillus nidulans. Gene 26:205–221
Tudzynski P, Esser K (1982) Extrachromosomal genetics of Cephalosporium acremonium: II. Development of a mitochondrial DNA hybrid vector replicating in Saccharomyces cerevisiae. Curr Genet 6:153–158
Tudzynski P, Esser K (1985) Mitochondrial DNA for gene cloning in eukaryotes. In: Bennet JW, Lasure LL (eds) Gene manipulations in fungi. Academic Press, New York, London, pp 403–416
Tudzynski P, Stahl U, Esser K (1982) Development of a eukaryotic cloning system in Podospora anserina. I. Long lived mutants as potential recipients. Curr Genet 6:219–222
Author information
Authors and Affiliations
Additional information
Present address: Institut für Biotechnologie, Fachgebiet Mikrobiologie, Techn. Universität Berlin, Seestr. 13, D-1000 Berlin 65
Rights and permissions
About this article
Cite this article
Stahl, U., Leitner, E. & Esser, K. Transformation of Penicillium chrysogenum by a vector containing a mitochondrial origin of replication. Appl Microbiol Biotechnol 26, 237–241 (1987). https://doi.org/10.1007/BF00286316
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00286316