Skip to main content
Log in

Recombinational stability of replicating plasmids in Aspergillus nidulans during transformation, vegetative growth and sexual reproduction

Current Genetics Aims and scope Submit manuscript

Abstract

Plasmids containing the AMA1 replicon are capable of autonomous maintenance in Aspergillus nidulans. It has been reported previously that these plasmids can form concatenates by recombination in a transformed mycelium, and up to 10% of molecules are involved in such events. The present study demonstrates that plasmid recombination, although frequent during transformation, rarely occurs during vegetative growth. As a result, the structure and phenotypic stability of AMA1 plasmids generally remains unaltered for many asexual (conidial) generations. It is also evident that plasmid replication does not require specific recombination events in the AMA1 palindrome. However, during sexual reproduction, autonomous plasmids exhibit increased recombination, which results in both plasmid concatenation and integration into the chromosome.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aleksenko AY (1994a) Co-integration of transforming DNAs in Aspergillus nidulans: a model using autonomously replicating plasmids. Curr Genet 26:352–358

    Google Scholar 

  • Aleksenko AY (1994b) Instant gene bank' method adapted for cloning a mutant benomyl-resistance allele in a replicative vector in Aspergillus nidulans. Fungal Genet Newslett 41:15–16

    Google Scholar 

  • Birnboim HL, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Blin N, Stafford DW (1976) A general method for isolation of highmolecular-weight DNA from eukaryotes. Nucleic Acids Res 3:2303–2308

    Google Scholar 

  • Bowyer P, Osbourn AE, Daniels MJ (1994) An „instant gene bank” method for heterologous gene cloning: complementation of two Aspergillus nidulans mutants with Gaeumannomyces graminis DNA. Mol Gen Genet 242:448–454

    Google Scholar 

  • Chung CT, Niemela SL, Miller RH (1988) One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci USA 86:2172–2176

    Google Scholar 

  • Clutterbuck AJ (1974) Aspergillus nidulans. In: King RC (ed) Handbook of genetics, vol 1. Bacteria, bacteriophage and fungi. Plenum Press, New York, pp 447–510

    Google Scholar 

  • Clutterbuck AJ, Gems DH, Robertson S (1994) The ARp1 Aspergillus replicating plasmid. In: Powell KA, Renwick A, Peberdy JF (eds) The genus Aspergillus from taxonomy and genetics to industrial application. Plenum Press, New York London, pp 189–196

    Google Scholar 

  • Durrens P, Green PM, Arst HN Jr, Scazzocchio C (1986) Heterologous insertion of transforming DNA and generation of new deletions letions associated with transformation in Aspergillus nidulans. Mol Gen Genet 203:544–549

    Google Scholar 

  • Futcher AB (1986) Copy number amplification of the 2 μm circle plasmid of Saccharomyces cereviseae. J Theor Biol 119:197–204

    Google Scholar 

  • Gems DH, Clutterbuck AJ (1993) Co-transformation with autonomously-replicating helper plasmids facilitates gene cloning from an aspergillus nidulans gene library. Curr Genet 24:520–524

    Google Scholar 

  • Gems D, Johnstone IL, Clutterbuck AJ (1991) An autonomously replicating plasmid transforms Aspergillus nidulans at high frequency. Gene 98:61–67

    Google Scholar 

  • Gems D, Aleksenko A, Belenky L, Robertson S, Ramsden M, Vinetski Y, Clutterbuck AJ (1994) An ‘instant gene bank’ method for gene cloning by mutant complementation. Mol Gen Genet 242:467–471

    Google Scholar 

  • Johnstone IL, Hughes SC, Clutterbuck AJ (1985) Cloning an Aspergillus nidulans developmental gene by transformation. EMBO J 4:1307–1311

    Google Scholar 

  • Käfer E (1961) The processes of spontaneous and induced somatic segregation in vegetative nuclei of Aspergillus nidulans. Genetics 46:1581–609

    Google Scholar 

  • Morrissey JA, Cashmore AM (1992) Use of cis-acting mutation to study the role of FLP-mediated recombination in the maintenance of native yeast 2 μm plasmids. Mol Microbiol 6:3101–3107

    Google Scholar 

  • Orr-Weaver TL, Szostak JW (1983) Multiple tandem plasmid integration in Saccharomyces cereviseae. Mol Cell Biol 3:747–749

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbour Laboratory. Cold Spring Harbor, New York

    Google Scholar 

  • Varma A, Kwon-Chung KJ (1994) Formation of a minichromosome in Cryptococcus neoformans as a result of electroporative transformation. Curr Genet 26:54–61

    Google Scholar 

  • Verdoes JC, Punt PJ, van den Berg P, Debets F, Stouthamer AH, van den Hondel CAMJJ (1994) Characterization of an efficient gene cloning strategy for Aspergillus niger based on an autonomously replicating plasmid: cloning of the nicB gene of A. niger. Gene 146:159–165

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H. T. Jacobs

Rights and permissions

Reprints and permissions

About this article

Cite this article

Aleksenko, A.Y., Clutterbuck, A.J. Recombinational stability of replicating plasmids in Aspergillus nidulans during transformation, vegetative growth and sexual reproduction. Curr Genet 28, 87–93 (1995). https://doi.org/10.1007/BF00311886

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00311886

Key words

Navigation