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  • 1
    Call number: 01-Personalabt:298 ; 01-Personalabt:300 ; 01-Personalabt:301 ; 01-Personalabt:303
    Keywords: Employee-management relations in government / Law and legislation / Germany
    Pages: xxxviii, 373 p.
    Edition: 11. Aufl.
    ISBN: 3-8073-1991-3
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    01-Personalabt:298 departmental collection or stack – please contact the library
    01-Personalabt:300 departmental collection or stack – please contact the library
    01-Personalabt:301 departmental collection or stack – please contact the library
    01-Personalabt:303 departmental collection or stack – please contact the library
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  • 2
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    Stuttgart : Thieme
    Call number: 03-E:35
    Keywords: Biochemistry ; Molecular Biology
    Pages: ix, 725 p. : ill.
    ISBN: 3-13-116681-9
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    03-E:35 departmental collection or stack – please contact the library
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  • 3
    Keywords: CANCER ; CELLS ; EXPRESSION ; GROWTH ; tumor ; TUMOR-CELLS ; CELL ; Germany ; MODEL ; screening ; DISEASE ; GENE ; GENES ; PROTEIN ; DRUG ; ACTIVATION ; INFECTION ; MECHANISM ; CARCINOGENESIS ; mechanisms ; SKIN ; BIOLOGY ; SUSCEPTIBILITY ; DISCOVERY ; virus ; TUMOR PROGRESSION ; genetics ; CANCER-CELLS ; ONCOGENE ; TRANSFORMATION ; REPLICATION ; INTERFERON ; HaCaT ; Ras ; CYTOTOXICITY ; GTPASE ; signaling ; ONCOLOGY ; HUMAN CANCER ; oncolytic virus ; NEWCASTLE-DISEASE-VIRUS ; DRUG DISCOVERY ; MOLECULAR-MECHANISMS ; Newcastle disease virus ; RAS SIGNALING PATHWAY ; oncolytic ; carcinogenesis model ; CELL BIOLOGY ; SIRNA ; Genetic ; INTERFERON RESPONSE ; Molecular mechanisms ; PERMISSIVENESS ; Rac1
    Abstract: Oncolytic Newcastle disease virus (NDV) replicates selectively in most human tumor cells but not in normal cells. The relationship between tumorigenesis and the selective susceptibility of most tumor cells to oncolytic NDV replication is poorly understood. A multistage skin carcinogenesis model derived from non-tumorigenic HaCaT cells was used to systematically investigate the molecular mechanisms involved in the oncolytic NDV-sensitivity associated with tumorigenic transformation. No significant differences in interferon signaling were observed between the virus-sensitive tumor cells and the virus-resistant non-tumorigenic parental cells. Oncogenic H-Ras, which had been used for tumorigenic transformation, was shown to be necessary for virus replication but was not sufficient to render cells susceptible to NDV replication. By using an siRNA screening approach to search for virus-sensitizing genes in the tumorigenic cells, we could identify the small GTPase Rac1 as an oncogenic protein that is essential for NDV replication and anchorage-independent growth in tumorigenic cells. Furthermore, Rac1 expression was sufficient to render non-tumorigenic cells susceptible to NDV replication and to oncolytic cytotoxicity. This study establishes Rac1 as a link between tumorigenesis and oncolytic virus sensitivity in the HaCaT multistage skin carcinogenesis model. Oncogene (2010) 29, 2205-2216; doi: 10.1038/onc.2009.507; published online 25 January 2010
    Type of Publication: Journal article published
    PubMed ID: 20101224
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  • 4
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The lysC/asd gene cluster of Corynebacterium glutamicum ATCC 13032 was cloned and sequenced. The lysC locus coding for aspartokinase consists of two in-frame overlapping genes, lysCα encoding a protein of 421 amino acids (Mr 44300) and lysCβ encoding a protein of 172 amino acids (Mr 18600). The C. glutamicum aspartokinase was purified and found to contain two proteins of Mr 47000 and Mr 18000.A C. glutamicum mutant expressing a feedback-resistant aspartokinase was shown to be changed in a single base pair of the lysCβ gene, leading to an amino acid exchange in the β-subunit of the aspartokinase. In addition, the identified mutation was found to be responsible for the enhanced expression of the asd gene located downstream of lysC.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-072X
    Keywords: Streptomyces ghanaensis ; Plasmid pSG2 ; Plasmid curing ; Shuttle plasmids ; Low copy broad host range Streptomyces plasmids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A plasmid, pSG2, was isolated from Streptomyces ghanaensis and characterized by electron microscopy, buoyant density measurement, and restriction enzyme analysis. The length of 13.8 kb, single restriction sites for HindIII, EcoRV and PvuII and the possibility of deleting non-essential regions of the plasmid made pSG2 a suitable basic replicon for vector development. pSG2 has a copy number of about four. Plasmid pSG2 was fused to a pACYC184 derivative modified to harbour a thiostrepton resistance gene. The resulting plasmid, designated pSW1, is a 16.6 kb shuttle plasmid which replicates in Escherichia coli and in several Streptomyces strains, including S. ghanaensis, S. lividans and S. viridochromogenes. Replacement of a Bg/II-fragment of plasmid pSG2 by a fragment encoding thiostrepton resistance resulted in a low copy 12.2 kb Streptomyces plasmid. This plasmid, designated pSW2, is a Streptomyces broad host range plasmid.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Streptomyces coelicolor “Müller” DSM3030 excretes a lysozyme comprising both β-1,4-N-acetyl-and β-1,4-N,6-O-diacetyl muramidase activities. The lysozyme is named Cellosyl. Gene libraries have been established using genomic DNA from the wild-type strain, S. coelicolor DSM3030, and from an overproducing mutant, S. coelicolor HP1, which exhibits about a twofold increase in lysozome production. The lysozyme-encoding genes (cel) from both strains were detected by oligodeoxynucleotide hybridization. The nucleotide sequence of the cel genes isolated from both strains was shown to be identical. The different levels of lysozyme production could not be correlated with any mutations at the cel gene locus. The cel gene isolated from the wild-type strain could not be expressed in some other species of Streptomyces. However, self-cloning of the cel gene into S. coelicolor DSM3030 and HP1 resulted in a 2.5-fold increase in lysozyme production.
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  • 7
    ISSN: 1420-9071
    Keywords: C4-dicarboxylate transport ; dct genes ; energy source ; regulation ; Rhizobium meliloti ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract TheRhizobium meliloti C4-dicarboxylate transport (Dct) system is essential for an effective symbiosis with alfalfa plants. C4-dicarboxylates are the major carbon source taken up by bacteroids. Genetic analysis of Dct− mutant strains led to the isolation of thedct carrier genedctA and the regulatory genesdctB anddctD. The carrier genedctA is regulated in free-living cells by the alternative sigma factor RpoN and the two-component regulatory system DctB/D. In addition, DctA is involved in its own regulation, possibly by interacting with DctB. In bacteroids, besides the DctB/DctD system an additional symbiotic activator is thought to be involved indctA expression. Further regulation ofdctA in the free-living state is reflected by diauxic growth of rhizobia, with succinate being the preferred carbon source. The tight coupling of C4-dicarboxylate transport and nitrogen fixation is revealed by a reduced level of C4-dicarboxylate transport in nitrogenase negative bacteroids.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The deliberate release of genetically engineered microorganisms requires a thorough characterization of the microbes in question. For the two bioluminescentRhizobium meliloti strains, L1 and L33 [Selbitschka et al. (1992) Mol Ecol 1: 9–19; Selbitschka et al. (1995) FEMS Microbiol Ecol 16:223–232], designated for field release, the sites of genetic modifications in the chromosomes were sequenced from amplified genomic DNA. This indicated no unexpected alterations in the nucleotide sequence. The bioluminescent phenotype was stably inherited over more than 100 generations in liquid cultures. The presence of the luciferase gene in both strains did not have secondary effects on a variety of metabolic pathways as assessed by the Biolog GN system. A specific polymerase chain reaction amplification, based on the chromosomal insertion site of theluc cassette, allowed the discrimination between the two strains and thus simplifies monitoring. The RecA-deficient strain L1 showed a strongly (more than 90%) reduced ability to nodulate alfalfa in competition with its parent strainR. meliloti 2011 and its RecA+ counterpart L33.
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  • 9
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Streptomyces coelicolor “Müller” is known to excrete the lysozyme N-acetylmuramidase. Culture filtrates of this strain form a characteristic halo on agar plates containing freeze-dried Micrococcus luteus cells (lysoplate technique). The halo consists of a clear inner zone and a turbid outer ring. Simulation experiments showed that the turbid outer ring is most probably produced by lysozyme whereas the clear inner zone can be considered to be due to an additional protease action. Using the lysoplate technique UV- and NTG-mutagenized strains of S. coelicolor “Müller” were screened for mutants defective in lysozyme production. Two mutants, SC11 and SC12, were identified. The mutant SC11 was selected for complementation studies. First, a transformation system was established. The use of a soft-agar overlay method was necessary to yield high regeneration rates of SC11 protoplasts. The plasmid vector pEB15 could be transferred into this mutant strain with an efficiency of 105 transformants/μg DNA. The high efficiency allowed shot-gun cloning experiments. Genomic DNA of S. coelicolor “Müller” digested with Sau3A and inserted into pEB15 was introduced into the mutant SC11. A complemented mutant was identified. A 2.9 kilobase pair (kb) DNA fragment was found which restored the lysozyme production of both mutants, SC11 and SC12. According to the diameter of the produced halos the complemented mutant SC11 was suggested to produce more lysozyme than the wildtype strain.
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  • 10
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Intact and osmotically sensitive cells of Corynebacterium glutamicum can be efficiently transformed by electroporation. This was shown by using the plasmid vector pUL-330 (5.2 kb), containing the kanamycin resistance gene of transposon Tn5. The following electric parameters yielded efficient transformation. For intact cells: one exponentially decaying field pulse $$E = E_0 exp( - t/\tau _E )$$ with time constants $$\tau _E = 450 - 500$$ and with initial field intensities of E 0=35–40 kV cm-1; prepulse temperature 20°C. Cell regeneration (survival) was 100%–80%. Transformation efficiency can be increased by an additional freeze and thaw cycle of the cells, prior to electroporation. Lysozyme treated cells (osmotically sensitive) were transformed with three successive pulses of E 0=25–30 kV cm-1. Cell regeneration under these conditions was found to be 20–30%. The optimum yield of transformants/μg plasmid-DNA was 3×103 for intact cells, 2×104 for intact cells which were frozen and thawed twice and 7×104 for osmotically sensitive cells if the cell suspension was pulsed at a cell density of 1–3×108/ml and at a DNA concentration of 0.2 μg/ml up to ≤2 μg/ml. The data obtained for osmotically sensitive cells suggest that the temperature increase accompanying the electric field pulse enhances colony formation and transformation efficiency if the initial prepulse temperature is ≥20°C, although regeneration of electroporated C. glutamicum cells starts to decrease at temperatures≥20°C.
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