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  • 1995-1999  (2)
  • CMV  (1)
  • nitrogen  (1)
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  • 1
    ISSN: 1573-4943
    Keywords: Cytomegalovirus proteinase ; CMV ; calorimetry ; ultracentrifugation ; mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Human cytomegalovirus (CMV) encodes a unique serine proteinase that is required in the maturation of the viral capsid. The CMV proteinase can undergo autocatalytic activation and is subject to proteolytic self-inactivation. Mutant enzyme forms were prepared to eliminate the initial autoprocessing site and thus form an active single-chain protein for structure-function studies. Two mutants of CMV proteinase were cloned and expressed inEscherichia coli. The A143V mutant was a conservative substitution at the first internal cleavage site. The S132A mutant modified one of the triad of residues responsible for catalytic activity. Through the use of computer-controlled high-cell-density fermentations the mutant proteins were expressed inE. coli at ∼170mg/L as both soluble (∼40% of total) and inclusion-body forms (∼60% of total). The soluble enzyme was purified by standard methods; inclusion-body protein was isolated by standard methods after refolding and solubilization in guanidine or urea. Sedimentation equilibrium and sedimentation velocity analyses reveal that the enzyme undergoes concentration-dependent aggregation. It exhibits a monomer⇔dimer equilibrium (K d =1ΜM) at low concentrations and remains dimeric at high concentrations (28 mg/ml). Differential scanning calorimetry data for protein thermal unfolding fit best to a non-two-state model with two components (T m =52.3 and 55.3
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5036
    Keywords: borescope ; cherry ; nitrogen ; persistence ; root ; season
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The persistence of white roots in cherry (Prunus avium) grown under two levels of nitrogen (high and low), was studied directly using a non-destructive borescope system. Plant growth and nitrogen uptake was increased by the extra nitrogen. Persistence times of white roots were significantly affected by several factors such as nitrogen level, time of appearance and depth in the pot. Suberisation of white roots tended to be late in the growing season, and roots produced in the middle of May persisted as white for significantly longer (5.9 weeks) than those produced in early July (2.5 weeks). Also, roots that appeared at the foot of the pot remained white for longer (5.6 weeks) than roots at the surface of the pot (2.7 weeks). Results also suggest that white root persistence was greater for trees with the lower rate of application of nitrogen (4.5 weeks under low nitrogen compared to 3.6 under high nitrogen).
    Type of Medium: Electronic Resource
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