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  • 1
    Keywords: Forschungsbericht
    Type of Medium: Online Resource
    Pages: 1 Online-Ressource (7 Seiten, 282,70 KB)
    Language: German
    Note: Engl. Berichtsbl. u.d.T.: de.NBI - conceptional phase - establishment of a preliminary administration office , Förderkennzeichen BMBF 031A421A , Unterschiede zwischen dem gedruckten Dokument und der elektronischen Ressource können nicht ausgeschlossen werden , Systemvoraussetzungen: Acrobat reader.
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  • 2
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The stringent response in Corynebacterium glutamicum was investigated. Sets of rrn–cat fusions were constructed in their native chromosomal position to examine the effects of amino acid starvation in a rel+ strain and a Δrel mutant defective in (p)ppGpp metabolism. The expression of the six rrn operons in the rel+ control was stringently regulated and reduced to 79% upon induction of amino acid starvation. The Δrel mutant displayed a relaxed regulation and was unable to reduce the rrn expression under amino acid depletion conditions. In addition, the Δrel mutant grew more slowly in minimal medium than a rel+ control. This growth effect was restored by a plasmid-encoded copy of rel or, alternatively, by supplementation of the minimal medium with the amino acid mixture casamino acids. In particular, the Δrel strain of C. glutamicum displayed a requirement for the amino acids histidine and serine.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The tetracycline resistance region of the 50-kb R-plasmid pTP10 from the clinical isolate Corynebacterium striatum M82B was analyzed in Corynebacterium glutamicum ATCC 13032 and confined to a 4.4-kb SphI-SalI DNA fragment. Nucleotide sequence analysis revealed two open reading frames, termed tetA and tetB, specifying proteins of 513 and 528 amino acids, respectively. The deduced amino acid sequences of tetAB displayed similarity to ATP-binding cassette transporters including StrV and StrW of Streptomyces glaucescens which are proposed to play a role in the export of streptomycin-like aminoglycosides. An antibiotic susceptibility screening in C. glutamicum showed that the tetAB genes confer resistance to tetracycline, oxytetracycline and to the structurally and functionally unrelated β-lactam antibiotic oxacillin.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In a recent proteomics study we have shown that the mcbR gene of Corynebacterium glutamicum ATCC 13032 most probably encodes a transcriptional repressor of the TetR type, which regulates the expression of at least six genes involved in the synthesis of sulphur-containing amino acids. By means of DNA microarray hybridizations we detected 86 genes with enhanced transcription in an mcbR mutant when compared with the wild-type strain. Bioinformatic analysis identified the inverted repeat 5′-TAGAC-N6-GTCTA-3′ as a consensus sequence within the upstream region of 22 genes and operons, suggesting that the transcription of at least 45  genes is directly controlled by the McbR repressor. These 45 genes encode a variety of functions in  (S-adenosyl)methionine  and  cysteine  biosynthesis, in sulphate reduction, in uptake and utilization of sulphur-containing  compounds  and  in  transcriptional regulation. The function of the inverted repeat motif as potential McbR binding site in front of the genes hom, cysI, cysK, metK and mcbR was verified experimentally by competitive electrophoretic mobility shift analysis. A systematic search for the potential effector substance modulating the function of McbR revealed that only S-adenosylhomocysteine prevented the binding of McbR to its target sequence. These results indicate that the transcriptional repressor McbR directly regulates a set of genes comprising all aspects of transport and metabolism of the macroelement sulphur in C. glutamicum. As the activity of McbR is modulated by S-adenosylhomocysteine, a major product of transmethylation reactions, the results point also to a novel regulatory mechanism in bacteria to control the biosynthesis of S-adenosylmethionine.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In a recent study, the putative regulatory gene cg0012 was shown to belong to the regulon of McbR, a global transcriptional regulator of sulphur metabolism in Corynebacterium glutamicum ATCC 13032. A deletion of cg0012, now designated ssuR (sulphonate sulphur utilization regulator), led to the mutant strain C. glutamicum DK100, which was shown to be blocked in the utilization of sulphonates as sulphur sources. According to DNA microarray hybridizations, transcription of the ssu and seu genes, encoding the sulphonate utilization system of C. glutamicum, was considerably decreased in C. glutamicum DK100 when compared with the wild-type strain. Electrophoretic mobility shift assays with purified SsuR protein demonstrated that the upstream regions of ssuI, seuABC, ssuD2 and ssuD1CBA contain SsuR binding sites. A nucleotide sequence alignment of the four DNA fragments containing the SsuR binding sites revealed a common 21 bp motif consisting of T-, GC- and A-rich domains. Mapping of the transcriptional start sites in front of ssuI, seuABC, ssuD2 and ssuD1CBA indicated that the SsuR binding sites are located directly upstream of identified promoter sequences and that the ssu genes are expressed by leaderless transcripts. Binding of the SsuR protein to its operator was shown to be diminished in vitro by the effector substance sulphate and its direct assimilation products adenosine 5′-phosphosulphate, sulphite and sulphide. Real-time reverse transcription polymerase chain reaction experiments verified that the expression of the ssu and seu genes was also repressed in vivo by the presence of sulphate or sulphite. Therefore, the regulatory protein SsuR activates the expression of the ssu and seu genes in C. glutamicum in the absence of the preferred sulphur source sulphate.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 123 (1994), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Efficient electroporation of Escherichia coli with plasmid DNA isolated from Corynebacterium glutamicum depends on the use of Mcr-deficient E. coli strains. The transformation frequency increased nearly 800-fold when the Mcr-deficient E. coli DH5αMCR was used instead of E. coli DH5α. We used E. coli strains with different mutations in the methyl-specific restriction systems to show that McrBC-deficiency is sufficient to generate this effect. The results imply that C. glutamicum DNA contains methylcytosine in specific sequences recognized by the E. coli McrBC system.
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  • 7
    ISSN: 1432-072X
    Keywords: Key wordsCorynebacterium glutamicum ; Amino acid ; production ; Branched-chain amino acids ; Isoleucine ; uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By complementation analysis of an isoleucine-uptake-deficient Escherichia coli strain, it was shown that a 1.6-kb HindIII-StuI fragment of Corynebacterium glutamicum ATCC 13032, located downstream of the aecD gene, encodes an isoleucine uptake system. Sequence analysis revealed that the complementing fragment carried an open reading frame, termed brnQ, that encodes a protein with sequence similarities to branched-chain amino acid carriers of gram-positive and gram-negative bacteria. The brnQ gene specifies a predominantly hydrophobic protein of 426 amino acid residues with a calculated molecular mass of 44.9 kDa. A topology prediction by neural network computer analysis suggests the existence of 12 hydrophobic segments that most probably form transmembrane α-helices. A C. glutamicum mutant strain harboring a defined deletion of brnQ in the chromosome showed a considerably lower isoleucine uptake rate of 0.04 nmol min–1 mg (dry mass)–1 as compared to the wild-type strain rate of 1.2 nmol min–1 mg (dry mass)–1. Overexpression of brnQ by means of a tac promotor resulted in an elevated uptake rate for isoleucine of 11.3 nmol min–1 mg (dry mass)–1. Evidently, the brnQ gene encodes the only transport system in C. glutamicum directing isoleucine uptake.
    Type of Medium: Electronic Resource
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  • 8
    Publication Date: 2023-06-21
    Description: 〈ns4:p〉The German Network for Bioinformatics Infrastructure (de.NBI) is a national and academic infrastructure funded by the German Federal Ministry of Education and Research (BMBF). The de.NBI provides (i) service, (ii) training, and (iii) cloud computing to users in life sciences research and biomedicine in Germany and Europe and (iv) fosters the cooperation of the German bioinformatics community with international network structures. The de.NBI members also run the German node (ELIXIR-DE) within the European ELIXIR infrastructure. The de.NBI / ELIXIR-DE training platform, also known as special interest group 3 (SIG 3) ‘Training & Education’, coordinates the bioinformatics training of de.NBI and the German ELIXIR node. The network provides a high-quality, coherent, timely, and impactful training program across its eight service centers. Life scientists learn how to handle and analyze biological big data more effectively by applying tools, standards and compute services provided by de.NBI. Since 2015, more than 300 training courses were carried out with about 6,000 participants and these courses received recommendation rates of almost 90% (status as of July 2020). In addition to face-to-face training courses, online training was introduced on the de.NBI website in 2016 and guidelines for the preparation of e-learning material were established in 2018. In 2016, ELIXIR-DE joined the ELIXIR training platform. Here, the de.NBI / ELIXIR-DE training platform collaborates with ELIXIR in training activities, advertising training courses via TeSS and discussions on the exchange of data for training events essential for quality assessment on both the technical and administrative levels. The de.NBI training program trained thousands of scientists from Germany and beyond in many different areas of bioinformatics.〈/ns4:p〉
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , NonPeerReviewed
    Format: application/pdf
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