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  • Chemical Engineering  (2)
  • Escherichia coli  (2)
  • 1990-1994  (4)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 891-906 
    ISSN: 0006-3592
    Keywords: ribosome vector ; cloned-gene expression ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An expression system utilizing specialized ribosomes has been constructed with β-galactosidase as the product. Ribosomes specific for lacZ mRNA are generated due to a mutation within the anti-Shine-Dalgarno region of a plasmidborne 16S rRNA gene that is complementary to a mutation within the ribosome-binding site of lacZ. Hence, a subpopulation of ribsomes specific for translation of the cloned gene mRNA is produced. Transcription of the lacZ gene is regulated by the tac promoter, while transcription of the mutated rrnB locus is controlled by the λPL promoter. Batch experiments indicate that full induction of both operons (2 mM IPTG, 42°C) leads to maximal β-galactosidase activity per cell at levels 35% higher than that obtained using a wild-type ribosome expression system. Using a novel, site-directed mutagenesis technique, construction of the specialized ribosome vector is outlined, and the results of lacZ expression are presented as transcription of both the cloned-gene and the specialized-ribosome locus are induced.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 38 (1991), S. 397-412 
    ISSN: 0006-3592
    Keywords: Escherichia coli ; protein synthesis ; metabolic limitations ; cloned-gene expression ; β-galactosidase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Earlier experiments in our lab investigated the metabolic limitations of cloned-gene expression in bacterial cells (for over-production of β-lactamase). These experiments showed that the steady-state concentration of ribosomal RNA decreased upon plasmid amplification while both the synthesis rate and steady-state β-lactamase mRNA level increased significantly. This appeared to indicate substantial limitation exist within the transnational machinery of the bacterial cell at high copy numbers. To establish the generality of this phenomenon, the impact increasing protein expression from pa plasmid by chemically inducing a strong promoter while maintaining constant copy number has been investigated. A plasmid has been constructed which contains the lacZ gene under control of the tac promoter and contains the parB stability locus to maintain plasmid stability. Using this vector, β-galactosidase expression in chemostat cultures operated at specific growth rates of 0.6 h-1 was induced with IPTG such that enzyme activity was varied over a 460-fold range. When fully induced β-galactosidase protein production represented 14 wt % of total cell protein. As transcription was induced, the synthesis rate of the β-galactosidase mRNA increased 42-fold while the steady-state level of β-galactosidase mRNA increased only fourfold. This indicates stability may play a larger role for β-galactosidase expression with a strong promoter than seen with β-lactamase production in the elevated copy number system. Furthermore, rRNA synthesis rates increased at high expression rates as seen in the copy number experiments. However, unlike the amplified-plasmid system, the steady-state levels of rRNA increased as well. Since the total protein levels closely followed the steady-state level of eRNA, transnational limitations are again suggested for the chemically induced transcription system.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Stamford, Conn. [u.a.] : Wiley-Blackwell
    Polymer Engineering and Science 32 (1992), S. 157-171 
    ISSN: 0032-3888
    Keywords: Chemistry ; Chemical Engineering
    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: Residual stress measurements and distortion analyses have been conducted on injection molded plaques made from polypropylene (PP) and a short glass-fiber filled polypropylene (GFPP). The residual stress analyses include measurements both parallel and perpendicular to the direction of flow during mold filling. Residual stresses are very anisotropic in GFPP, but not in PP. The residual stress levels in PP fall on aging at room temperature, whereas in GFPP the proportion of stress relaxation is smaller, and significant stresses remain even after heating to elevated temperatures. A significant contribution to distortion has been linked to the ejection process, and the long- and short-term distortion of moldings is discussed within the framework of the properties of the materials measured here.
    Additional Material: 19 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    Stamford, Conn. [u.a.] : Wiley-Blackwell
    Polymer Engineering and Science 33 (1993), S. 1390-1399 
    ISSN: 0032-3888
    Keywords: Chemistry ; Chemical Engineering
    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: A test program was developed to compare the viscoelastic behavior of an untoughened epoxy (R922) with a toughened epoxy (R6376). Creep/recovery tests were performed on neat resin as well as [ ± 40° ]2s composites using T500 or IM7 graphite fibers. The composite specimens were subjected to at least five cycles of load/unload at each stress level. The goal was to study the viscoelastic response of a flexibilized and unflexibilized epoxy and to try and observe any microstructural changes that might occur. Neat resin tests did not show a significant difference in behavior between the two systems. Single cycle behavior was found to differ significantly from multiple cycle behavior. Recovery strains were observed to be much higher than predicted from linear theory. Linear parameters were also seen to vary with stress level; however, when a plasticity-based approach was used to model the creep behavior, the same parameters could then be used at all load levels. Creep and recovery behavior were seen to differ significantly. This observation leads to support for the theory of Schapery (1) that creep and recovery should be considered as different behavior in contrast to the assumptions of linear and nonlinear theory. In addition, the large difference between neat resin and composite data suggests that micromechanics models, which rely on constituent properties, will lead to large errors if neat resin properties are used to characterize composite viscoelastic behavior. Different fibers showed no influence on results. Scanning electron microscopy failed to reveal any microstructural changes.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
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