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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 29 (1995), S. 50-57 
    ISSN: 1432-0983
    Keywords: Triticale/wheat ; atpA/atp9 genes ; Mitochondrial mRNA processing ; Nucleo-cytoplasmic interactions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene region coding for subunits α and 9 of the mitochondrial ATP synthase exhibit an identical DNA sequence in wheat, rye, and the intergeneric hybrid triticale (xTriticosecale Wittmack). However, co-transcripts containing both genes show different sizes depending on the nuclear genotype. To investigate nuclear-mitochondrial interactions leading to this variation, we performed a comparative transcript analysis with various lines carrying defined nuclear and cytoplasmic genotypes. Northern analyses showed that all wheat lines investigated possess a single atpA/atp9 mRNA of 2.6 kb, whereas in rye and five independent triticale lines an additional transcript of 2.35 kb appeared. Primer-extension and RNase-protection analyses indicate that the co-transcripts of this gene have staggered 5′ termini in some lines, whereas the 3′ termini seem to be similar in wheat, rye, and triticale. Transcription is initiated at position-338/-339 upstream of the atpA gene in all lines investigated, giving rise to a 2.6-kb mRNA. In rye and triticale, staggered 5′ termini were observed closer to the translational start. The DNA sequences upstream of these termini exhibit homology to plant mitochondrial-processing sites, therefore the proximal 5′ ends are most probably generated by RNA processing. As the processing event occurs more frequently in triticale carrying the Triticum timopheevi cytoplasm, trans-acting factors from rye are likely to interact with other cytoplasmic factors resulting in the observed RNA modification. Most interestingly, the T. timopheevi cytoplasm inducing male sterility in alloplasmic wheat, fails to generate the CMS phenotype in triticale. The data support our hypothesis that nuclear factors affect mitochondrial gene expression and thus control sexual fertility in wheat and triticale.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Sordaria macrospora ; DNA transformation ; pcbC Promoter ; Heterologous integration ; Pulsed-field gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ascomycete Sordaria macrospora was transformed using different plasmid molecules containing the bacterial hygromycin B resistance gene (hph) under the control of different expression signals. The highest transformation frequency was obtained with vector pMW1. On this plasmid molecule, expression of the hph gene is directed by the upstream region of the isopenicillin N synthetase gene (pcbC) from the deuteromycete Acremonium chrysogenum. Southern analysis suggests that the vector copies are integrated as tandem repeats into the S. macrospora chromosomes and that duplicated sequences are most probably not inactivated by methylation during meiosis. Furthermore, the hygromycin B resistance (hygR) is not correlated with the number of integrated vector molecules. Electrophoretic karyotyping was used to further characterize S. marcospora transformants. Five chromosomal bands were separated by pulsed-field gel electrophoresis (PFGE) representing seven chromosomes with a total genome size of 39.5 Mb. Hybridization analysis revealed ectopic integration of vector DNA into different chromosomes. In a few transformants, major rearrangements were detected. Transformants were sexually propagated to analyze the fate of the heterologous vector DNA. Although the hygR phenotype is stably maintained during mitosis, about a third of all lines tested showed loss of the resistance marker gene after meiosis. However, as was concluded from electrophoretic karyotyping, the resistant spores showed a Mendelian segregation of the integrated vector molecules in at least three consecutive generations. Our data indicate that heterologous marker genes can be used for transformation tagging, or the molecular mapping of chromosomal loci in S. macrospora
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 3 (1981), S. 151-156 
    ISSN: 1432-0983
    Keywords: Podospora anserina plDNA ; Integral part of mtDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary As previously reported, a ccc DNA with a contour length of 0.75 µm and molecular weight of 2.4 kb (termed plasmid-like, p1DNA) is the causative agent of senescence in the fungus Podospora anserina. Its postulated location in mtDNA was proved correct by the following experiments: 1. Restriction analysis of mtDNA resulted in different molecular weights for both, juvenile (95 kb) and senescent (30 kb) mtDNA. The construction of a detailed restriction map made evident the fact that senescent mtDNA comprises only a part of its juvenile counterpart. 2. Hybridization experiments (Southern blots) between 3H-labelled plDNA and mtDNA cleaved by restriction juvenile mtDNA are homologous to plDNA. 3. Fine mapping experiments (construction of restriction maps and heteroduplex experiments) between plDNA integrated into a bacterial vector and its postulated equivalent, derived from juvenile mtDNA and also integrated into a bacterial vector, allowed a precise determination of the site of plDNA insertion within the juvenile mtDNA. All of these data fit into a previously published model in which, during aging, plDNA is excised from mtDNA and becomes autonomous for replication and function.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0983
    Keywords: Key wordsChlamydomonas reinhardtii ; Chlorophyll-a/b-binding protein ; Light regulation ; Plastid signal ; Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Chlamydomonas reinhardtii, expression of the lhcb1 gene encoding a chlorophyll-a/b-binding protein of photosystem II is highly regulated by light, inhibitors of chlorophyll synthesis, as well as by circadian rhythms. In light/dark synchronized cultures, the rapid increase of lhcb1 mRNA levels during the light phase is regulated primarily at the transcriptional level. We have used the arylsulphatase (ars) reporter gene to analyze the lhcb1 5′ upstream sequences for the presence of light-responsive elements. In transformants carrying chimeric reporter genes, accumulation of lhcb1/ars mRNA is markedly stimulated by light, with a time course similar to that of transcripts from the endogenous lhcb1 gene. Promoter deletion studies revealed that a 255-bp fragment of the lhcb1 5′ upstream region is sufficient to confer proper light regulation on the promoterless ars gene. Moreover, the region between positions –255 and –122 with respect to the start site of translation were found to contain one or more light-responsive elements. Strikingly, these sequences also seem to be involved in chloroplast-dependent lhcb1 gene expression as indicated by Northern analyses of transformants with photo-oxidatively damaged chloroplasts. This suggests that both light- and chloroplast-dependent expression of the lhcb1 gene are mediated by the same cis-acting elements.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0983
    Keywords: Keywords ATP citrate lyase ; Filamentous fungi ; Gene cluster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract ATP citrate lyase (ACL) catalyzes the formation of cytosolic acetyl-CoA, which is mainly used for the biosynthesis of fatty acids and sterols. In this paper, we show for the first time that in filamentous fungi two different subunits of ACL are encoded by two separate genes. This is in contrast to animals where ACL is encoded by a single gene. Data are presented on acl genes from the filamentous fungi Sordaria macrospora and Gibberella pulicaris. In S. macrospora, both genes, acl1 and acl2, are clustered within a region of 10 kb and are divergently transcribed.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 24 (1993), S. 421-427 
    ISSN: 1432-0983
    Keywords: Acremonium chrysogenum ; pcbC gene ; Transformation ; Gene disruption ; Pulsed-field gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cephalosporin C-producing fungus Acremonium chrysogenum was transformed to hygromycin B resistance using different vector constructs. These constructs contain sequences of the pcbC gene from A. chrysogenum, encoding isopenicillin N synthetase. Detailed analysis of transformants, including pulsed-field gel electrophoresis (PFGE), suggests that integration of multiple vector copies takes place predominantly via non-homologous integration. By increasing the length of vector-DNA homologous to genomic DNA, integration occurs more frequently into chromosome VI, carrying the endogencous pcbC gene copy. In gene disruption experiments, the length of vector homology required to obtain cephalosporin C-minus transformants was investigated. Inactivation of the pcbC gene was observed only when homologous fragments of more than 3.0 kb were used on both sites of the resistance cassette. Southern analysis indicated homologous, as well as heterologous, integration of recombinant DNA. The integration of multiple vector copies leads to the appearance of truncated pcbC transcripts.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0983
    Keywords: Podospora anserina ; Rearrangements of mtDNA ; Senescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mapping and transcription studies have revealed that in Podospora anserina the causative agent of senescence, a mitochondrial plasmid (p1DNA), is identical with intronl of the discontinuous gene for cytochrome-c-oxidase subunit 1 (COI), which is 2 kpb from the discontinuous gene for cytochrome b (Cytb). A mitochondrial mutant (ex1) devoid of the COI, but not of the Cytb gene provides longevity. A molecular model for the onset of senescence is presented.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 56 (2005), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Fungal morphogenesis and secondary metabolism are frequently associated; however, the molecular determinants connecting both processes remain largely undefined. Here we demonstrate that CPCR1 (cephalosporin C regulator 1 from Acremonium chrysogenum), a member of the winged helix/regulator factor X (RFX) transcription factor family that regulates cephalosporin C biosynthesis, also controls morphological development in the β-lactam producer A. chrysogenum. The use of a disruption strain, multicopy strains as well as several recombinant control strains revealed that CPCR1 is required for hyphal fragmentation, and thus the formation of arthrospores. In a ΔcpcR1 disruption strain that exhibits only hyphal growth, the wild-type cpcR1 gene was able to restore arthrospore formation; a phenomenon not observed for ΔcpcR1 derivatives or non-related genes. The intracellular expression of cpcR1, and control genes (pcbC, egfp) was determined by in vivo monitoring of fluorescent protein fusions. Further, the role of the forkhead transcription factor AcFKH1, which directly interacts with CPCR1, was studied by generating an Acfkh1 knockout strain. In contrast to CPCR1, AcFKH1 is not directly involved in the fragmentation of hyphae. Instead, the presence of AcFKH1 seems to be necessary for CPCR1 function in A. chrysogenum morphogenesis, as overexpression of a functional cpcR1 gene in a ΔAcfkh1 background has no effect on arthrospore formation. Moreover, strains lacking Acfkh1 exhibit defects in cell separation, indicating an involvement of the forkhead transcription factor in mycelial growth of A. chrysogenum. Our data offer the potential to control fungal growth in biotechnical processes that require defined morphological stages for optimal production yields.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 25 (1994), S. 34-40 
    ISSN: 1432-0983
    Keywords: Acremonium chrysogenum ; β-tubulin gene ; Homologous transformation system ; Benomyl resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Theβ−tubulin gene was isolated from the filamentous fungusAcremonium chrysogenum using a heterologous gene probe to screen anA. chrysogenum lambda library. Sequencing of theA. chrysogenum gene revealed a mosaic gene which contains five exons and four intervening sequences. The exons encode for a polypeptide of 447 amino-acid residues which showed a high degree of similarity when compared with amino-acid sequences from β-tubulins of other eukaryotes. The introns are characterized by typical consensus sequences found in intervening sequences from other filamentous fungi. In-vitro mutagenesis of codon 167 of the β-tubulin gene resulted in the substitution of a phenylalanine by a tyrosine in the corresponding polypeptide sequence. The mutated gene was used successfully in the transformation and co-transformation ofA. chrysogenum to benomyl resistance. The molecular analysis of transformants provided evidence that they contain the mutated β-tubulin gene in addition to the wild-type gene, as was proved by Southern-hybridization analysis and direct sequencing of PCR amplification products.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 19 (1991), S. 73-76 
    ISSN: 1432-0983
    Keywords: Acremonium (Cephalosporium) species ; Cephalosporin C producer ; Electrophoretic karyotype
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A restriction fragment length polymorphism (RFLP) analysis was performed on six related Acremonium strains. With respect to the restriction fragment pattern, all strains of A. chrysogenum were indistinguishable from each other but showed distinctive differences from those of A. strictum, A. flavum and Cephalosporium polyvaleurum. Using pulsed-field gel electrophoresis, we obtained different chromosome patterns from most of the Acremonium strains. Remarkably, the pattern varies in three related A. chrysogenum strains which also differ in their rate of cephalosporin C biosynthesis. The electrophoretic karyotyping was confirmed by the location of rDNA genes on separate chromosomes. Our data indicate that chromosome translocations in industrial strains may be responsible for increased β-lactam synthesis.
    Type of Medium: Electronic Resource
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