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  • Articles  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 27 (1979), S. 247-253 
    ISSN: 1432-0827
    Keywords: Osteoblast ; Osteoclast ; Osteoprogenitor cells ; Fracture ; Chimera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Previous studies have shown that differences in nuclear morphology are generally sufficient to determine the species origin of cells in interspecific grafts between the Japanese quail and domestic chicken. Most quail nuclei possess 1–3 large nucleolus-associated masses of heterochromatin. Chick cells, on the other hand, usually present a more diffuse, stippled distribution of nuclear heterochromatin. Quail embryonic limb rudiments, some with and some without established marrow cavities, were explanted and grown on the chorioallantoic membrane of the chick. Three to five days post-grafting, the explants were surgically fractured and allowed to heal. Tissues were collected and histologically processed during the latter period. The fractures healed completely within 5–6 days and no callus was established in the process. The nuclear staining pattern of the osteoblasts and osteocytes throughout the rudiments and at the fracture site indicated that they were derived from the graft. Possible sources for these cells included the periosteum, endosteum, and posthypertrophy chondrocytes. By contrast, most of the nuclei in the osteoclasts were chick-like and were apparently derived from cells originating in the host. Because the quail-like heterochromatin marker was normally present in a small number (2.5%) of chick osteoclast nuclei and was lacking in about 5% of native quail osteoclast nuclei, the precise extent of the participation of donor, i.e., quail bone and marrow stromal cells in osteoclast formation, could not be determined. However, the data suggest that in large measure the precursor cells for most osteoclasts were hematogenously derived and were carried to the grafted rudiments by the blood vascular system.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5028
    Keywords: cyanogenic glucosides ; cytochrome P450 ; E. coli expression ; oxime reconstitution ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA encoding the multifunctional cytochrome P450, CYP71E1, involved in the biosynthesis of the cyanogenic glucoside dhurrin from Sorghum bicolor (L.) Moench was isolated. A PCR approach based on three consensus sequences of A-type cytochromes P450 – (V/I)KEX(L/F)R, FXPERF, and PFGXGRRXCXG – was applied. Three novel cytochromes P450 (CYP71E1, CYP98, and CYP99) in addition to a PCR fragment encoding sorghum cinnamic acid 4-hydroxylase were obtained. Reconstitution experiments with recombinant CYP71E1 heterologously expressed in Escherichia coli and sorghum NADPH–cytochrome P450–reductase in L-α-dilaurylphosphatidyl choline micelles identified CYP71E1 as the cytochrome P450 that catalyses the conversion of p-hydroxyphenylacetaldoxime to p-hydroxymandelonitrile in dhurrin biosynthesis. In accordance to the proposed pathway for dhurrin biosynthesis CYP71E1 catalyses the dehydration of the oxime to the corresponding nitrile, followed by a C-hydroxylation of the nitrile to produce p-hydroxymandelonitrile. In vivo administration of oxime to E. coli cells results in the accumulation of the nitrile, which indicates that the flavodoxin/flavodoxin reductase system in E. coli is only able to support CYP71E1 in the dehydration reaction, and not in the subsequent C-hydroxylation reaction. CYP79 catalyses the conversion of tyrosine to p-hydroxyphenylacetaldoxime, the first committed step in the biosynthesis of the cyanogenic glucoside dhurrin. Reconstitution of both CYP79 and CYP71E1 in combination with sorghum NADPH-cytochrome P450–reductase resulted in the conversion of tyrosine to p-hydroxymandelonitrile, i.e. the membranous part of the biosynthetic pathway of the cyanogenic glucoside dhurrin. Isolation of the cDNA for CYP71E1 together with the previously isolated cDNA for CYP79 provide important tools necessary for tissue-specific regulation of cyanogenic glucoside levels in plants to optimize food safety and pest resistance.
    Type of Medium: Electronic Resource
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