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  • 2000-2004  (4)
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  • 1
    Electronic Resource
    Electronic Resource
    Development of a novel quadruple auxotrophic host transformation system by argB gene disruption using adeA gene and exploiting adenine auxotrophy in Aspergillus oryzae (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 239 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We previously designed a triple auxotrophic host-vector system in Aspergillus oryzae by isolating red-colored adenine auxotrophic mutants upon UV mutagenesis of a double auxotrophic host (niaD−sC−). In the present study an effort to exploit this system and construct a novel quadruple auxotrophic host was made by disrupting the argB gene involved in arginine biosynthesis. The argB gene-disruption cassette was generated by fusion PCR, which required only two steps of PCR to insert the selectable marker, adeA, into the target argB gene. The chimeric DNA fragment was transformed into the triple auxotrophic strain (niaD−sC−adeA−) and the argB disruptants were obtained with a high rate of efficiency (approximately 40%). The argB disruptants were characterized by normal colony color and reversal of arginine auxotrophy by introduction of the wild-type argB gene. Quadruple auxotrophic strains (niaD−sC−ΔargB adeA− or niaD−sC−ΔargB adeB−) were subsequently isolated upon UV mutagenesis of the triple auxotrophic strain (niaD−sC−ΔargB) followed by screening of red-colored colonies for adenine auxotrophy. The results obtained showed that the adeA gene served as an efficient selection marker in developing a novel host-vector system with quadruple auxotrophy in A. oryzae, thus providing a powerful tool to breed multiple auxotrophic mutants from a deuteromycete wherein sexual crossing is impossible.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.08.025
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    BaM/Pt multilayered films with fine grains and large perpendicular magnetic anisotropy for high density recording media (2001)
    [S.l.] : American Institute of Physics (AIP)
    Journal of Applied Physics 89 (2001), S. 6837-6839 
    Details
    ISSN: 1089-7550
    Source: AIP Digital Archive
    Topics: Physics
    Notes: A magnetoplumbite type of Ba ferrite (BaM) thin layer was deposited on a 9 nm thick Pt underlayer, and excellent c-axis orientation was observed even for an 8 nm thick BaM layer, which corresponds to only three or four BaM unit cells. The grain size was almost in the same range of 60–85 nm even when the BaM layer thickness tBaM decreased from 60 to 17 nm, and tBaM should be reduced below 10 nm to make a grain size smaller than 50 nm. However, the perpendicular coercivity Hc⊥ and squareness S⊥ decreased drastically from 2.6 to 0.5 kOe and from 0.6 to 0.2, respectively, with the decrease of tBaM from 60 to 8 nm because of higher demagnetizing field and susceptibility to thermal fluctuation. On the other hand, the [BaM(5–24 nm)/Pt(9 nm)]3 multilayer exhibited higher Hc⊥ and larger S⊥ than the BaM/Pt bilayer for the same BaM layer thickness and Hc⊥ and S⊥ of the [BaM(8 nm)/Pt(9 nm)]3 multilayer was 2.0 kOe and 0.6, respectively. It was clarified that the deposition of the BaM/Pt multilayer was very effective for achieving a high perpendicular magnetic anisotropy constant even for the ultrathin BaM layer. © 2001 American Institute of Physics.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1063/1.1359468
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Mouse-related dermatosis (2004)
    Oxford, UK : Blackwell Science Ltd
    International journal of dermatology 43 (2004), S. 0 
    Details
    ISSN: 1365-4632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-4632.2004.02299.x
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  • 4
    Electronic Resource
    Electronic Resource
    A HU-like gene mutation in Rhizobium leguminosarum bv. viciae affects the expression of nodulation genes (2004)
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 51 (2004), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: NodD is the major regulator of nod genes expression in rhizobia. Previously, a HU-like protein in Rhizobium leguminosarum bv. viciae has been identified to bind specifically with nod promoters and be involved in in vitro nodD transcription, but its in vivo function remained unknown. In this work we have cloned and sequenced the R. leguminosarum bv. viciae gene, named hurL, for this HU-like protein. Using the E. coli-expressed HurL proteins, we proved that HurL had high affinity to several nod promoters and showed a stimulation effect on in vitro nodD transcription at appropriate concentration. The R. leguminosarum bv. viciae hurL gene was mutated by insertion of a kanamycin resistance cassette. The obtained hurL mutant strain M704 exhibited poor growth under free-living conditions and failed to induce nodules on Pisum sativum cv. Frisson and Vicia hirsuta. Further studies of NodD production and nod genes-lacZ fusions expression in the hurL mutant revealed that inactivation of hurL led to severe impairment in the nodD expression, repression in the inducible expression of nodA and nodF, and slight enhancement in the expression of px2, a gene identified earlier in this lab. These results suggested that hurL might be required for maintaining the normal expression of nod genes in R. leguminosarum bv. viciae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.2003.03873.x
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