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  • 1
    Electronic Resource
    Electronic Resource
    Transformation of Aspergillus parasiticus using autonomously replicating plasmids from Aspergillus nidulans (1994)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 124 (1994), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract A genetic transformation system for the aflatoxin-producing fungus Aspergillus parasiticus using two autonomously replicating plasmids from A. nidulans (ARp1 and pDHG25) is reported. Transformation frequencies using the plasmid pDHG25 were from 5 × 102 to 2.5 × 104 transformants per 106 viable protoplasts and μg DNA. The stability of the plasmids in the transformants was also studied. This transformation system offers a new opportunity to clone genes related to aflatoxin production using appropriate aflatoxin-defective mutants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1994.tb07258.x
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  • 2
    Electronic Resource
    Electronic Resource
    Genetic maps of eight linkage groups of Aspergillus niger based on mitotic mapping (1993)
    Springer
    Current genetics 23 (1993), S. 47-53 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus niger ; Genetic maps ; Mitotic mapping ; AmdS transformants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper provides a genetic map of Aspergillus niger. At present 84 markers have been assigned to eight linkage groups. The chromosomal location of 60 markers is presented in this paper. The allocation of markers is based on recombination due to mitotic crossing over. Various methods for selection and analysis of homozygous recombinants were applied, using colour, auxotrophic and resistance markers. In addition, transformants carrying the heterologous Aspergillus nidulans gene coding for acetamidase (amdS) were used for mitotic mapping of markers in several linkage groups. In most of the transformants the amdS insert appeared to be centromere-distal to all known genetic markers, thus extending the eight linkage groups has been determined. On the basis of these and earlier experiments tentative genetic markers were found on both arms of the chromosomes, except for chromosomes II and IV. The genetic distance between markers and the centromere varies from about 10-4 (LG I, II, V) up to more than 10-2 (LGIII, VI, VIII). The total frequency of mitotic recombination per genome in this fungus has been estimated to be at least 1.2×10-1.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00336749
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  • 3
    Electronic Resource
    Electronic Resource
    Arginine and proline genes ofAspergillus niger (1992)
    Springer
    Antonie van Leeuwenhoek 61 (1992), S. 259-264 
    Details
    ISSN: 1572-9699
    Keywords: arginine mutants ; Aspergillus niger ; proline mutants ; linkage groups ; somatic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Aspergillus niger mutants defective in arginine or proline biosynthesis have been isolated and 12 genetic loci were identified. Mutation was induced by low doses UV, and mutants were isolated after filtration enrichment. The mutants were classified according to their phenotype in growth tests and were further characterized in complementation tests. The arginine auxotrophic mutants represent nine complementation groups. Three additional complementation groups were found for mutants that could grow on proline (two of them on arginine too). Linkage group analysis was done in somatic diploids obtained from a mutant and a master strain with genetic markers on six chromosomes. Thearg genes belong to six different linkage groups and thepro genes to two. Onearg-mutant could be complemented by transformation with theA. nidulans arg B + gene, and thisA. niger gene thus appeared to be homologous to theA. nidulans arg B. We isolated anA. niger strain with theargB gene tightly linked with thenicA1 marker. This strain is very suitable as acceptor for transformation with anargB-plasmid, because transformants with inserts on the homologous site can be recognized and analyzed genetically using thenicA1 marker gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00713933
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  • 4
    Electronic Resource
    Electronic Resource
    Isolation and molecular characterisation of the benzoate-para-hydroxylase gene (bphA) of Aspergillus niger: A member of a new gene family of the cytochrome P450 superfamily (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 192-197 
    Details
    ISSN: 1617-4623
    Keywords: Monooxygenase ; Recombinant DNA ; Overexpression ; Upstream reading frames ; Complementation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The gene coding for benzoate-para-hydroxylase (bphA) of Aspergillus niger was cloned using differential hybridisation techniques and complementation of mutants deficient in this enzyme activity. The nucleotide sequence of the gene was determined, the presence of two introns was shown and the transcription start and termination sites were determined. The structure of the mRNA upstream from the long open reading frame (ORF) is unusual. It contains two small, overlapping ORFs whose function is unknown. Comparison of the deduced amino acid sequence of the protein with the sequences present in the databanks, indicated a significant similarity of BPH to the superfamily of cytochrome P450 enzymes. Further analysis revealed that this protein is a member of a new P450 gene family designated P450LIII. The gene is designated CYP53. To increase the BPH activity of A. niger, multiple copies of the bphA gene were introduced into the genome of a recipient strain by transformation. Although increased intracellular levels of the BPH protein could be detected, the BPH enzyme activity was decreased, suggesting titration of another essential component.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00265053
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  • 5
    Electronic Resource
    Electronic Resource
    An electrophoretic karyotype of Aspergillus niger (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 264-268 
    Details
    ISSN: 1617-4623
    Keywords: Pulsed field gel electrophoresis ; Aspergillus niger ; Genome size ; amdS transformants ; Chromosomal size markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An electrophoretic karyotype of Aspergillus niger was obtained using contour-clamped homogeneous electric field (CHEF) gel electrophoresis. Chromosomesized DNA was separated into four bands. Seven of the eight linkage groups could be correlated with specific chromosomal bands. For this purpose DNA preparations from seven transformant strains of A. niger each carrying the heterologous amdS gene of Aspergillus nidulans on a different chromosome were analysed. Some of the assignments were confirmed with linkage groupspecific A. niger probes. The estimated sizes of the A. niger chromosome range from 3.5 to 6.6 Mb, based on gel migration relative to the chromosomes of Schizosaccharomyces pombe strains, Saccharomyces cerevisiae and A. nidulans. The total genome size of A. niger significantly exceeds that of A. nidulans and is estimated to be about 35.5–38.5 Mb. Electrophoretic karyotyping was used to allocate non-mutant rRNA genes and to estimate the number of plasmids integrated in a high copy number transformant.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00271560
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  • 6
    Electronic Resource
    Electronic Resource
    Genetic analysis of Aspergillus niger: Isolation of chlorate resistance mutants, their use in mitotic mapping and evidence for an eighth linkage group (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 453-458 
    Details
    ISSN: 1617-4623
    Keywords: Chlorate resistance ; Aspergillus niger ; Linkage groups ; Transformation ; Recombinant frequency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper describes the use of chlorate resistant mutants in genetic analysis of Aspergillus niger. The isolated mutants could be divided into three phenotypic classes on the basis of nitrogen utilization. These were designated nia, nir and cnx as for Aspergillus nidulans. All mutations were recessive to their wild-type allele in heterokaryons as well as in heterozygous diploids. The mutations belong to nine different complementation groups. In addition a complex overlapping complementation group was found. Evidence for the existence of eight linkage groups was obtained. Two linked chlorate resistance mutations and two tryptophan auxotrophic markers, which were unlinked to any of the known markers (Goosen et al. 1989), form linkage group VIII. We used the chlorate resistance mutations as genetic markers for the improvement of the mitotic linkage map of A. niger. We determined the linear order of three markers in linkage group VI as well as the position of the centromere by means of direct selection of homozygous cnxA1 recombinants. In heterozygous diploid cultures diploid chlorate resistant segregants appeared among conidiospores with a frequency of 3.9×10−2 (cnxG13 in linkage group I) to 2.1 × 10−2 (cnxD6 in linkage group 111). The mean frequency of haploid chlorate resistant segregants was 1.3 × 10−3. The niaD1 and niaD2 mutations were also complemented by transformation with the A. niger niaD + gene cloned by Unkles et al. (1989). Mitotic stability of ten Nia+ transformants was determined. Two distinct stability classes were found, showing revertant frequencies of 5.0 × 10−3 and 2.0 × 10−5 respectively.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00259411
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  • 7
    Electronic Resource
    Electronic Resource
    Genetic analysis ofamdS transformants ofAspergillus niger and their use in chromosome mapping (1990)
    Springer
    Molecular genetics and genomics 222 (1990), S. 284-290 
    Details
    ISSN: 1617-4623
    Keywords: AmdS transformants ; Aspergillus niger ; Mitotic mapping ; Recombination ; Mitotic stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary TheAspergillus nidulans gene coding for acetamidase (amdS) was introduced intoA. niger by transformation. Twelve Amd+ transformants were analysed genetically. TheamdS inserts were located in seven different linkage groups. In each transformant the plasmid was integrated in only a single chromosome. Our (non-transformed)A. niger strains do not grow on acetamide and are more resistant to fluoroacetamide than the transformants. Diploids hemizygous for theamdS insert have the Amd+ phenotype. We exploited the opportunity for two-way selection inA. niger: transformants can be isolated based on the Amd+ phenotype, whereas counter-selection can be performed using resistance to fluoroacetamide. On this basis we studied the phenotypic stability of the heterologousamdS gene inA. niger transformants as well as in diploids. Furthermore, we mapped the plasmid insert of transformant AT1 to the right arm of chromosome VI betweenpabA1 andcnxA1, providing evidence for a single transformational insert. The results also show that theamdS transformants ofA. niger can be used to localize non-selectable recessive markers and that the method meets the prerequisites for efficient mitotic mapping. We suggest the use ofamdS transformants for mitotic gene mapping in other fungi.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00633830
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