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  • 1
    Electronic Resource
    Electronic Resource
    Purification of macrolide 2′-phosphotransferase fromStreptomyces coelicolor Müller (1990)
    Springer
    Journal of industrial microbiology and biotechnology 6 (1990), S. 295-297 
    Details
    ISSN: 1476-5535
    Keywords: Phosphorylation ; Oleandomycin ; Macrolide 2′-phosphotransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary An enzyme that catalyzes 2′-O-phosphorylation of oleandomycin and several other macrolide antibiotics has been purified approximately 47-fold from cell-free extracts ofStreptomyces coelicolor Müller, NRRL 3532 (UC™ 5240). The reaction product was verified as being oleandomycin-2′-O-phosphate by mass spectrometry. As a result of purification, the enzyme was separated from two lincosaminide inactivating enzyme activities also present in the cell-free extract.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01575877
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  • 2
    Electronic Resource
    Electronic Resource
    Discrimination of closely homologous human genomic and viral sequences in cells and tissues: further characterization of Tmt (1994)
    Springer
    Journal of molecular histology 26 (1994), S. 545-552 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The experimentally derived parameter Tmt (tissue Tm) was defined previously to describe the end-point used for evaluation of the stringency of non-isotopic in situ hybridization and was found to differ from the theoretical melting temperature (Tm) for several HPV types. In this paper, the reasons for this discrepancy were investigated by performing a series of experiments with a variety of probes for both human genomic and integrated viral sequences in isolated and cultured normal and abnormal cells in addition to paraffin-embedded material. Tmt was shown to be dependent on several parameters of probe and target, and on the sensitivity of the detection system used but was not affected by aldehyde fixation or paraffin wax embedding under optimal conditions of nucleic acid unmasking. These data support the hypothesis that differences between Tmt and Tm may be due to the use of a different end-point for in situ hybridization analysis rather than biochemical alteration of DNA-DNA interactions in intact cells. Appropriate stringency conditions should therefore be determined by experiment rather than calculated theoretically for gene evaluation in cells and tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00158588
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  • 3
    Electronic Resource
    Electronic Resource
    Discrimination of closely homologous HPV types by nonisotopicin situ hybridization: definition and derivation of tissue melting temperatures (1990)
    Springer
    Journal of molecular histology 22 (1990), S. 545-554 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It is generally assumed that nucleic acid association duringin situ hybridization reactions is similar to that of nucleic acid association in solution. This assumption has been investigated by detecting closely homologous human papillomavirus types 6 and 11 byin situ hybridization as a model for the evaluation of stringency conditions in clinical biopsies. By examining matched and mismatched, labelled and target sequences under various stringency conditions, empirical DNA-DNA stability curves and their derivative equations for tissue melting temperatures (Tmt) were derived. The corresponding values for Tmt are 10–20°C higher than their solution equivalents. These data, supported by polymerase chain reaction experiments, demonstrate that closely homologous viral DNAs cross linked in tissue by formaldehyde fixation do not interact with the corresponding labelled probes as predicted from solution kinetic equations. This not only has theoretical implications but is also relevant to the accuracy of clinical diagnostic testing.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01005977
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  • 4
    Electronic Resource
    Electronic Resource
    The Pharmacokinetics of Trilostane and Ketotrilostane in an Interconverting System in the Rat (1992)
    Springer
    Pharmaceutical research 9 (1992), S. 464-468 
    Details
    ISSN: 1573-904X
    Keywords: trilostane ; ketotrilostane ; reversible metabolism ; pharmacokinetics ; metabolic interconversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The pharmacokinetics of trilostane and one of its metabolites ketotrilostane are described and characterized in the rat following the separate intravenous administration of trilostane and ketotrilostane. It was noted during these studies that the parent compound and its metabolite undergo metabolic interconversion–trilostane producing ketotrilostane and ketotrilostane generating trilostane. This result means that trilostane is conserved in the body by interconversion-being metabolized to ketotrilostane and then subsequently back to the “parent” drug, trilostane.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1015828027060
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  • 5
    Electronic Resource
    Electronic Resource
    Homologous recombination between plasmid DNA molecules in maize protoplasts (1991)
    Springer
    Molecular genetics and genomics 230 (1991), S. 209-218 
    Details
    ISSN: 1617-4623
    Keywords: Homologous recombination ; Protoplast transformation ; β-Glucuronidase ; Maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The requirements for homologous recombination between plasmid DNA molecules have been studied using the PEG (polyethylene glycol)-mediated transformation system of maize (Zea mays L.) protoplasts coupled with the transient expression assay for β-glucuronidase (GUS). Two plasmids were introduced into maize protoplasts; one plasmid (pB×26) contained a genomic clone of the Adh1 maize gene; the other plasmid (piGUS) was a promoterless construction containing part of intron A of the Adhl gene fused to the gusA coding sequence. Thus, the two vectors shared an effective homologous region consisting of a 459 by (Hindlll—PvuII) fragment of the yAdh1 intron A sequence. An active gusA fusion gene would result upon homologous recombination between the plasmids within the intron A sequence, and indeed GUS activity was observed in extracts following co-transformation of maize protoplasts with the two plasmids. The presence of recombinant DNA molecules in protoplast DNA isolated 1 day after co-transformation was verified using polymerase chain reactions (PCR) and Southern blots. For efficient homologous recombination, both plasmids had to be linearized. The recombination reaction was induced by restriction of the plasmid molecules either inside the effective homologous region or at the borders of the intron sequence. However, the presence of even small, terminal, nonhomologous sequences at the 3′ end of the pB×26 fragment inhibited the recombination reaction. Also, both ends of the linearized piGUS DNA molecules were involved in the recombination reaction. The results revealed some features of homologous recombination reactions occurring in plant cells which cannot be accommodated by mechanisms postulated for similar reactions in animal system and in lower eukaryotes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00290670
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  • 6
    Electronic Resource
    Electronic Resource
    The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction (1993)
    Springer
    Journal of molecular histology 25 (1993), S. 191-198 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The parameter Tmt has been defined by non-isotopic in situ hybridization and describes the tissue melting temperature (Tmt) of human papillomavirus (HPV) DNA sequences. In this study, multiple in situ hybridization signals for HPV types 16, 31 and 33 in individual archival biopsies hybridized with genomic probes are shown by polymerase chain reactions to be due to cross-hybridization of probe sequences to a single tissue target. Tmt is independent of viral type but depends on the homology between probe and target when using nick-translated whole genomic probes. The difference between Tm and Tmt is not due to the presence of viral capsid protein. Multiple HPV signals in archival material should not therefore be interpreted as indicative of multiple HPV infection unless adequate stringency conditions have been employed or they are present in morphologically distinct areas of the biopsy. Furthermore, extrapolation of calculated DNA homologies to non-isotopic in situ hybridization analysis may not be appropriate. A hybridization signal does not imply probe and target identity: this has implications for HPV typing in clinical material.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00163814
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