DNA-mediated transformation of the basidiomycete Coprinus cinereus.

Binninger, David; Skrzynia, Cécile; Pukkila, Patricia J.; Casselton, Lorna A.

doi:10.1002/j.1460-2075.1987.tb04828.x

Cited by 232 publications

(144 citation statements)

References 39 publications

Supporting

Mentioning

138

Contrasting

Unclassified

Order By: Relevance

“…In five transformants (no. 4,5,15,17,19), the ade2 probe hybridized to a 9.4-kb fragment in addition to many novel fragments, implicating ectopic integration. In these transformants, the SK probe hybridized to a majority of these novel fragments but not to the 9.4-kb VOL.…”

Section: Resultsmentioning

confidence: 96%

Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA

et al. 1993

View full text Add to dashboard Cite

A transformation scheme for Cryptococcus neoformans to yield high-frequency, integrative events was developed. Adenine auxotrophs from a clinical isolate of C. neoformans serotype A were complemented by the cryptococcal phosphoribosylaminoimidazole carboxylase gene (ade2) with a biolistic DNA delivery system.Comparison of two DNA delivery systems (electroporation versus a biolistic system) showed notable differences. The biolistic system did not require linear vectors and transformed each auxotrophic strain at similar fr-equencies. Examination of randomly selected transformants by biolistics showed that 15 to 40%O were stable, depending on the recipient auxotroph, with integrative events identified in all stable transformants by DNA analysis. Although the ade2 cDNA copy transformed at a low frequency, DNA analysis found homologous recombination in each of these transformants. DNA analysis of stable transformants receiving genomic ade2 revealed ectopic integration in a majority of cases, but approximately a quarter of the transformants showed homologous recombination with vector integration or gene replacement. This system has the potential for targeted gene disruption, and its efficiency will also allow for screening of DNA libraries within C. neoformans. Further molecular strategies to study the pathobiology of this pathogenic yeast are now possible with this transformation system.Cryptococcus neoformans is a pathogenic yeast with an unexplained predilection for infecting the central nervous system. There has been a significant increase in the number of human infections with this yeast over the last decade as the immunocompromised population has enlarged because of AIDS. Although epidemiological studies suggest that the increase in cryptococcal infection is due to more-susceptible hosts rather than increased virulence, the pathobiology of the organism needs further investigation. Several phenotypic factors associated with virulence have been identified for C. neoformans: the polysaccharide capsule (10,16,20), melanin production (19,20,26), and growth at 370C (20,26 Recent studies of C neofonnans have established basic molecular techniques for performing site-directed mutagenesis (9, 31). A C. neoformans uraS auxotrophic strain (serotype D) was recently reported to be transformed at a low efficiency by complementation with a cloned uraS gene delivered by electroporation. However, the number of integrative events was low and generally nonhomologous (9, 31). A total of 80 to 90% of the transformants were unstable and apparently contained extrachromosomal DNA, consisting of sequences of rearranged uraS-vector DNA with the addition of cryptococcal genomic DNA (8,9,31

show abstract

Section: Resultsmentioning

confidence: 96%

Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA

et al. 1993

View full text Add to dashboard Cite

show abstract

“…Expression of C. disseminatus genes in C. cinerea: C. cinerea monokaryon 218 (A3, B1, trp1.1,16, bad; Binninger et al 1987;Kü es et al 2002) was transformed according to the protocol given by Granado et al (1997). The trp1 1 vector pCc1001 (Binninger et al 1987) was used in cotransformations with plasmid pCR2.1-TOPO containing the entire Aa sublocus from strains TJ00/99.1 and TJ01/16.3 (plasmids 99.1Aa and 16.3Aa) and the entire Ab sublocus from strains TJ00/99.1 and TJ00/89.2 (99.1Ab and 89.2Ab).…”

Section: Methodsmentioning

confidence: 99%

“…The trp1 1 vector pCc1001 (Binninger et al 1987) was used in cotransformations with plasmid pCR2.1-TOPO containing the entire Aa sublocus from strains TJ00/99.1 and TJ01/16.3 (plasmids 99.1Aa and 16.3Aa) and the entire Ab sublocus from strains TJ00/99.1 and TJ00/89.2 (99.1Ab and 89.2Ab). Also used in cotransformation with pCc1001 were C25.B2.5 containing the two putative pheromone receptor genes CDSTE3.1 and CDSTE3.3 and three pheromone genes CDPHB1, CDPHB2.1, and CDPHB2.2 and C25_e1.10, a 10-kb subclone of C25.B2.5 containing CDSTE3.1 and CDPHB1 in vector pZERO (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

Evolution of the Bipolar Mating System of the Mushroom Coprinellus disseminatus From Its Tetrapolar Ancestors Involves Loss of Mating-Type-Specific Pheromone Receptor Function

et al. 2006

View full text Add to dashboard Cite

Mating incompatibility in mushroom fungi is controlled by the mating-type loci. In tetrapolar species, two unlinked mating-type loci exist (A and B), whereas in bipolar species there is only one locus. The A and B mating-type loci encode homeodomain transcription factors and pheromones and pheromone receptors, respectively. Most mushroom species have a tetrapolar mating system, but numerous transitions to bipolar mating systems have occurred. Here we determined the genes controlling mating type in the bipolar mushroom Coprinellus disseminatus. Through positional cloning and degenerate PCR, we sequenced both the transcription factor and pheromone receptor mating-type gene homologs from C. disseminatus. Only the transcription factor genes segregate with mating type, discounting the hypothesis of genetic linkage between the A and B mating-type loci as the causal origin of bipolar mating behavior. The mating-type locus of C. disseminatus is similar to the A mating-type locus of the model species Coprinopsis cinerea and encodes two tightly linked pairs of homeodomain transcription factor genes. When transformed into C. cinerea, the C. disseminatus A and B homologs elicited sexual reactions like native matingtype genes. Although mating type in C. disseminatus is controlled by only the transcription factor genes, cellular functions appear to be conserved for both groups of genes. M ATING in fungi is controlled by the loci that determine the mating type of an individual, and only individuals with differing mating types can mate. Basidiomycete fungi have evolved a unique mating system, termed tetrapolar or bifactorial incompatibility, in which mating type is determined by two unlinked loci; compatibility at both loci is required for mating to occur. The origin of the tetrapolar mating system in the basidiomycetes is likely to be ancient since it is observed in at least two of the three major lineages, the Ustilaginomycetes, or smut fungi, and the Hymenomycetes, primarily the mushroom fungi (Burnett 1975). Also unique to the basidiomycetes is the presence of multiple alleles at the mating-type loci that allows most individuals within a population to be mating compatible with one another. Only the mushroom-forming homobasidiomycetes possess large allelic series at both loci, typically termed the A and B mating-type loci (Whitehouse 1949;Raper 1966).The multiallelic tetrapolar mating system is considered to be a novel innovation that could have only evolved once (Raper 1966;Raper and Flexer 1971). For this reason, the ancestor of the homobasidiomycetes is accepted as having a tetrapolar mating system. Although most (65%) of the homobasidiomycetes possess a tetrapolar mating system, many species (25%) instead have a bipolar system controlled by a single locus with multiple alleles (Raper 1966). The distribution of bipolar species is scattered throughout the homobasidiomycete phylogeny, and bipolar species appear to have multiple independent origins from tetrapolar mating systems (Hibbett and Donoghue 2001). The popul...

show abstract

“…Groups of 12 cosmid clones were cultured on a plate of Luria broth/ ampicilline solid medium, mixed, and subjected to mini prep. Pooled DNAs were used to transform strain T140 (A3B1 trp1-1,1-6 dst1-1) as described by Binninger et al (1987). Transformants were cultured on minimal medium to purify the transformed mycelium and then mated to R1428F 1 #72 (A8B7 dst1-2).…”

Section: Methodsmentioning

confidence: 99%

The dst1 Gene Involved in Mushroom Photomorphogenesis of Coprinus cinereus Encodes a Putative Photoreceptor for Blue Light

et al. 2005

View full text Add to dashboard Cite

The homobasidiomycete Coprinus cinereus exhibits remarkable photomorphogenesis during fruiting-body development. Under proper light conditions, fruiting-body primordia proceed to the maturation phase in which basidia in the pileus undergo meiosis, producing sexual spores, followed by stipe elongation and pileus expansion for efficient dispersal of the spores. In the continuous darkness, however, the primordia do not proceed to the maturation phase but are etiolated: the pileus and stipe tissues at the upper part of the primordium remain rudimentary and the basal part of the primordium elongates, producing ''dark stipe.'' In this study we genetically analyzed five strains that produce dark stipes even if light conditions promoting the maturation are given and then characterized one of them, Uar801 (dst1-1). The dst1 gene was cloned as a DNA fragment that rescues the dst1-1 mutation. Dst1 is predicted to be a protein of 1175 amino acids that contains two PAS domains, a coiled-coil structure, and a putative, glutamine-rich, transcriptional activation domain (AD). One of the PAS domains exhibits significant similarity to the LOV domains of known bluelight receptors, suggesting that Dst1 is a blue-light receptor of C. cinereus. The dst1-1 mutation is predicted to truncate the putative AD in the C-terminal region.

show abstract

DNA-mediated transformation of the basidiomycete Coprinus cinereus.

Cited by 232 publications

References 39 publications

Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA

Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA

Evolution of the Bipolar Mating System of the Mushroom Coprinellus disseminatus From Its Tetrapolar Ancestors Involves Loss of Mating-Type-Specific Pheromone Receptor Function

The dst1 Gene Involved in Mushroom Photomorphogenesis of Coprinus cinereus Encodes a Putative Photoreceptor for Blue Light

Contact Info

Product

Resources

About