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  • 1
    ISSN: 1617-4623
    Keywords: Schizosaccharomyces pombe ; DNA-damage inducibility ; Damage-responsive element ; Upstream activating sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract TheSchizosaccharomyces pombe rhp51 + gene encodes a recombinational repair protein that shares significant sequence identities with the bacterial RecA and theSaccharomyces cerevisiae RAD51 protein. Levels ofrhp51 + mRNA increase following several types of DNA damage or inhibition of DNA synthesis. Anrhp51::ura4 fusion gene was used to identify the cis-acting promoter elements involved in regulatingrhp51 + expression in response to DNA damage. Two elements, designated DRE1 and DRE2 (fordamage-responsiveelement), match a decamer consensus URS (upstream repressing sequence) found in the promoters of many other DNA repair and metabolism genes fromS. cerevisiae. However, our results show that DRE1 and DRE2 each function as a UAS (upstream activating sequence) rather than a URS and are also required for DNA-damage inducibility of the gene. A 20-bp fragment located downstream of both DRE1 and DRE2 is responsible for URS function. The DRE1 and DRE2 elements cross-competed for binding to two proteins of 45 and 59 kDa. DNase I footprint analysis suggests that DRE1 and DRE2 bind to the same DNA-binding proteins. These results suggest that the DRE-binding proteins may play an important role in the DNA-damage inducibility ofrhp51 + expression.
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  • 2
    ISSN: 1617-4623
    Keywords: Gβ protein ; Schizosaccharomyces pombe ; Sporulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract ASchizosaccharomyces pombe homolog of mammalian genes encoding G proteinβ subunits,gpb1 +, was cloned by the polymerase chain reaction using primer pairs that correspond to sequences conserved in several Gβ genes of other species followed by screening of genomic and cDNA libraries. Thegpb1 gene encodes 317 amino acids that show 47% homology with human Gβ 1 and Gβ 2 and 40% homology withSaccharomyces cerevisiae Gβ protein. Disruption of thegpb1 gene indicated that this gene is not required for vegetative cell growth. However,gpb1-disrupted haploid cells mated and sporulated faster than wild-type cells, both in sporulation (MEA) and in complex medium (YE): when examined 23 h after transfer to sporulation medium, 35% ofgpb1-disrupted haploid pairs had undergone conjugation and sporulation, whereas only 3–5% of wild-type haploid pairs had done so. Overexpression of thegpb1 gene suppressed this facilitated conjugation and sporulation phenotype ofgpb1-disrupted cells but did not cause any obvious effect in wild-type cells. Co-disruption of one of the twoS. pombe Gα-subunit genes,gpa2, in thegpb1-disrupted cells did not change the accelerated conjugation and sporulation phenotype of thegpb1 − cells. However, co-disruption of theras1 gene abolished thegpb1 − phenotype. These results suggest that Gpbl is a negative regulator of conjugation and sporulation that apparently works upstream of Ras1 function inS. pombe. The possible relationship of Gpbl to two previously identified, putative Gα proteins ofS. pombe is discussed.
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  • 3
    ISSN: 1617-4623
    Keywords: Key words Gβ protein ; Schizosaccharomyces pombe ; Sporulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Schizosaccharomyces pombe homolog of mammalian genes encoding G protein β subunits, gpb1 +, was cloned by the polymerase chain reaction using primer pairs that correspond to sequences conserved in several Gβ genes of other species followed by screening of genomic and cDNA libraries. The gpb1 gene encodes 317 amino acids that show 47% homology with human Gβ1 and Gβ2 and 40% homology with Saccharomyces cerevisiae Gβ protein. Disruption of the gpb1 gene indicated that this gene is not required for vegetative cell growth. However, gpb1-disrupted haploid cells mated and sporulated faster than wild-type cells, both in sporulation (MEA) and in complex medium (YE): when examined 23 h after transfer to sporulation medium, 35% of gpb1-disrupted haploid pairs had undergone conjugation and sporulation, whereas only 3–5% of wild-type haploid pairs had done so. Overexpression of the gpb1 gene suppressed this facilitated conjugation and sporulation phenotype of gpb1-disrupted cells but did not cause any obvious effect in wild-type cells. Co-disruption of one of the two S. pombe Gα-subunit genes, gpa2, in the gpb1-disrupted cells did not change the accelerated conjugation and sporulation phenotype of the gpb1 − cells. However, co-disruption of the ras1 gene abolished the gpb1 − phenotype. These results suggest that Gpbl1 is a nebative regulator of conjugation and sporulation that apparantely works upstream of Ras1 function in S. pombe. The possible relationship of Gpb1 to two previously indentified, putative Gα proteins of S. pombe is discussed.
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  • 4
    ISSN: 0935-9648
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Physics
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 0935-6304
    Keywords: Cyclic carbonates ; thermal stability ; chemical stability ; gas phase chromatography ; liquid phase chromatography ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: ---No abstract
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1040-7685
    Keywords: microcolumn liquid chromatography ; liquid chromatography/mass spectrometry ; temperature programming ; optimization methods ; fatty acids ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A temperature-programmed microcolumn liquid chromatography/mass spectrometry (LC/MS) system has been developed as a practical alternative to conventional solvent-programmed LC/MS. Good reproducibility of solute retention was achieved in a specially modified oven that permitted linear temperature programming between 40 and 100°C at rates of 0.1 to 0.5°C min-1. Through theoretical modelling studies performed under isocratic and isothermal conditions, the optimum separation under temperature-programmed conditions was rapidly and accurately predicted. This method enabled the complete resolution of saturated and unsaturated fatty acids in a fish oil dietary supplement with identification from their characteristic fragmentation patterns under electron-impact MS conditions. The mass spectra from the LC/MS system correlated very well with those from a standard reference library to facilitate spectral searching methods.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0887-3585
    Keywords: crystals ; bacterial esterase ; X-ray diffraction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Large crystals of arylesterase from Pseudomonas fluorescens have been grown at room temperature using ammonium sulfate as a precipitant. They grow to dimensions of 0.7 × 0.7 × 0.6 mm3 within a month. The crystals belong to the trigonal space group P31 (or P32), with unit cell dimensions of a= 147.12 Å and c= 131.08 Å. The asymmetric unit seems to contain six molecules of dimeric aryles-terase, with corresponding crystal volume per protein mass (VM) of 2.53 Å3/Da and solvent fraction of 51.5% by volume. The crystals diffract to at least 2.2 Å Bragg spacing when exposed to X-rays from a rotating-anode source. X-ray data have been collected to 2.9 Å Bragg spacing from native crystals. © 1993 Wiley-Liss, Inc.
    Additional Material: 1 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    ISSN: 0538-8066
    Keywords: Chemistry ; Physical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A vacuum ultraviolet photolysis of C2H5Br at 147 nm was studied over a pressure range of 0.5-50 torr at 298 K. The effects of additives He and NO were also investigated.The principal reaction products were found to be C2H4 and C2H6, with lesser yields of CH4 and C2H2. With increasing pressure the product quantum yields Φi of C2H4, CH4, and CH2H6 remained constant, while that of C2H2 decreased from 0.03 to almost 0. The effect of He as an additive was found to be extremely small on the quantum yields of the major products. Addition of NO completely suppresses the formation of CH4, C2H2, and C2H6, and reduces partially the production of C2H4. The primary processes appear to involve two electronically excited states. One state mainly yields C2H4 by molecular elimination of HBr and is thought to be due to a Rydberg transition. The other state decomposes to C2H5 and Br radicals by C—Br bond fission. These two competitive reaction modes contribute to the photodecomposition in proportions of 50% and 50%. The extinction coefficient for C2H5Br at 147 nm and at 298 K has been determined as ∊ = (1/PL) In(Io/It) = 712 ± 7 atm-1 · cm-1.
    Additional Material: 2 Ill.
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  • 9
    ISSN: 0959-8103
    Keywords: poly(esterurethane) ; copolyamide segments ; thermal and mechanical properties ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Physics
    Notes: Thermoplastic polyurethane elastomers (TPUs) from diol-terminated poly(ethylene adipate) (PEA), 1,4-butanediol (BD) and 4,4′-diphenylmethane-diisocyanate (MDI) were modified by copolymerizing with diamine-terminated nylon-6/6,6 copolyamide (CPA) oligomers. The effects of content and molecular weight of CPA segments on the thermal and mechanical properties of TPU were studied. PEA segments showed enhanced crystallization when some of the hard segments were replaced by CPA segments, showing weaker CPA-PEA interaction. The crystallinity of the hard segments was reduced, probably due to some interaction and phase mixing between hard and CPA segments. The modulus of TPU also decreased, more markedly with CPA segments of higher molecular weight.
    Additional Material: 7 Ill.
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  • 10
    ISSN: 1042-7147
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Small-angle neutron scattering studies have been used to clarify several aspects of the internal structure of latexes and subsequent film formation modes. This paper reviews work both around the world and at Lehigh University on those subjects. Two points have been made clear: (1) The appearance of core-shell phenomena in latexes depends on the size of the polymer chain to that of the latex particle; the phenomenon is most marked when the radius of gyration of the chain is about one fifth as big as the latex radius. (2) Strength build-up during film formation depends on the extent of interdiffusion of the chains. For moderate molecular weights, interdiffusion distances of one radius of gyration yield maximum strength. For both moderate (250,000 g/mol) and high (2,000,000 g/mol) molecular weights, full strength was achieved in two hour's annealing time at 144°C.
    Additional Material: 11 Ill.
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