Summary
To develop a transformation system in the filamentous fungus Podospora anserina we have selected ura5 mutants deficient in orotidylic acid pyrophosphorylase using a positive screening. These mutants could be transformed to prototrophy by an hybrid vector carrying the ura5 gene of this organism. The properties of the transformants have been analysed. In most cases integration of the transforming vector occurred outside the ura5 locus and frequently repeated tandem copies of the vector were found. Reversion of the transformants could also be selected and we found that it can occur by exact or only partial excision of the integrated vector.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- bp:
-
base pairs
- kb:
-
1,000 bp
- EtBr:
-
ethidium bromide
- PEG:
-
polyethyleneglycol
References
Balance DJ, Turner G (1985) Gene 36:321–331
Bégueret J, Razanamparany V, Perrot M, Barreau C (1984) Gene 32:487–492
Bernet J (1965) Ann Sci Nat Bot 6:611:768 Bernet J (1965) Ann Sci Nat Bot 6:611:768
Birnboim HC, Doly J (1979) Nucleic Acids Res 7:1513–1523
Boeke JD, Lacroute F, Fink GR (1984) Gen Genet 197:345–346
Brygoo Y, Debuchy R (1985) Mol Gen Genet 200:128–131
Case MF, Schweizer M, Kushner SR, Giles H (1979) Proc Natl Acad Sci USA 76:5259–5263
Clewell DB, Helinski DR (1972) J Bacteriol 110:1135–1146
Esser K (1974) Podospora anserina. In: King RC (ed) Handbook of genetics. Plenum Press, New York London, p 513
Hanahan D (1983) J Mol Biol 166:557–580
Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933
Kinsey JA, Rambosek JA (1984) Mol Cell Biol 4:117–122
Rigby PWJ, Diekmann M, Rhodes C, Berg P (1977) J Mol Biol 113:237–251
Rizet G, Engelmann C (1949) Rev Cytol Biol Veg 11:201–304
Sakaguchi I, Yamamoto M (1982) Proc Natl Acad Sci USA 79:7819–7823
Southern EM (1975) J Mol Biol 98:503–517
Stiles JI (1983) Methods Enzymol 101:290–300
Tilburn J, Scazzochio C, Taylor GC, Zabicky-Zisslman JH, Lockington RA, Davis RW (1983) Gene 26:205–221
Wernars K, Goosen T, Wennekes LMJ, Visser J, Bos CJ, van den Broek HWJ, van Gorcon RFM, van den Hondel CAMJJ, Pouwels PH (1985) Curr Genet 9:361–368
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Razanamparany, V., Bégueret, J. Positive screening and transformation of ura5 mutants in the fungus Podospora anserina: characterization of the transformants. Curr Genet 10, 811–817 (1986). https://doi.org/10.1007/BF00418527
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00418527