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J E Landegent N Jansen in de Wal G J van Ommen F Baas J J de Vijlder P van Duijn M Van der Ploeg 《Nature》1985,317(6033):175-177
During the past few years, several methods have been developed for the detection of specific nucleic acid sequences by in situ hybridization using non-radioactive labels such as fluorochromes, cytochemically detectable enzymes and electron-dense markers. These methods are preferable to autoradiography in terms of speed of performance and topological resolution. Their limited sensitivity, however, has so far restricted their use to the detection of repeated sequences. Here we report single gene detection with a procedure using 2-acetylaminofluorene (AAF)-modified probes, immunoperoxidase cytochemistry and reflection-contrast microscopy. We confirmed the autoradiographic data on the localization of the human thyroglobulin (Tg) gene to the distal end of the long arm of chromosome 8. A mixture of cosmid cHT2-derived subclones of the 3' part of the Tg gene, 22.3 kilobase pairs (kbp) in total, was used as a hybridization probe. This procedure can be used to map other unique sequences, if genomic clones are available from which clones with an appropriate amount of inserts can be isolated. 相似文献
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Neurotransmission from mossy fibre terminals onto cerebellar granule cells is almost certainly mediated by L-glutamate. By taking advantage of the small soma size, limited number of processes and short dendrite length of granule cells, we have obtained high-resolution recordings of spontaneous miniature excitatory postsynaptic currents (m.e.p.s.cs) and evoked currents in thin cerebellar slices. Miniature currents have a similar time-course and pharmacology to evoked currents and consist of an exceptionally fast non-NMDA (N-methyl-D-aspartate) component (measured rise-time, 200 microseconds; estimated pre-filtered rise-time less than 100 microseconds; decay time constant, tau = 1.0 ms), followed by 50 pS NMDA channel openings that are directly resolvable. We could find no evidence for the recent proposal that miniature currents in granule cells are mediated solely by NMDA channels with a novel time course. The non-NMDA receptor component of m.e.p.s.cs has a skewed amplitude distribution, which suggests potential complications for quantal analysis. The difference in time course between the m.e.p.s.cs reported here and other synaptic currents in the brain could reflect differences in synaptic function or electrotonic filtering; the relative contribution of these possibilities has yet to be established. 相似文献
117.
Descartes is always concerned about knowledge. However, the Galileo affair in 1633, the reactions to his Discourse on method, and later his need to reply to objections to his Meditations provoked crises in Descartes’s intellectual development the import of which has not been sufficiently recognized. These events are the major reasons why Descartes’s philosophical position concerning how we know and what we may know is radically different at the end of his life from what it was when he began. We call this later position Descartes’s epistemic stance and contrast it with his earlier methodological, metaphysical realism. Yet Descartes’s epistemic views cannot be separated from other aspects of his work, for example, his views concerning God, causality, metaphysics, and the nature of science. A further meta-implication is that serious errors await any scholar who cites early Cartesian texts in support of late Cartesian positions, or who uses later texts in conjunction with early ones to support a reading of Descartes’s philosophy. 相似文献
118.
We have studied the chemistry of aminoacyl AMP to model reactions at the 3' terminus of aminoacyl tRNA for the purpose of understanding the origin of protein synthesis. The present studies relate to the D, L preference in the esterification of 5'-AMP. All N-acetyl amino acids we studied showed faster reaction of the D-isomer, with a generally decreasing preference for D-isomer as the hydrophobicity of the amino acid decreased. The beta-branched amino acids, Ile and Val, showed an extreme preference for D-isomer. Ac-Leu, the gamma-branched amino acid, showed a slightly low D/L ratio relative to its hydrophobicity. The molecular basis for these preferences for D-isomer is understandable in the light of our previous studies and seems to be due to preferential hydrophobic interaction of the D-isomer with adenine. The preference for hydrophobic D-amino acids can be decreased by addition of an organic solvent to the reaction medium. Conversely, peptidylation with Ac-PhePhe shows a preference for the LL isomer over the DD isomer. 相似文献
119.
J. -M. Weber 《Cellular and molecular life sciences : CMLS》1992,48(6):557-564
The study of metabolic fuel provision and its regulation has reached an exciting stage where specific molecular events can be correlated with parameters of the organism's ecology. This paper examines substrate supply pathways from storage sites to locomotory muscle mitochondria and discusses ecological implications of the limits for maximal flux through these pathways. The relative importance of the different oxidative fuels is shown to depend on aerobic capacity. Very aerobic, endurance-adapted animals such as long distance migrants favor the use of lipids and intramuscular fuels over carbohydrates and circulatory fuels. The hypothesis of functional co-adaptation between oxygen and metabolic fuel supply systems allows us to predict that the capacity of several biochemical processes should be scaled with maximal oxygen consumption. Key enzymes, transmembrane transporter proteins, glucose precursor supply and soluble fatty acid transport proteins must all be geared to support higher maximal glucose and fatty acid fluxes in aerobic than in sedentary species. 相似文献
120.
J A Martignetti A A Aqeel W A Sewairi C E Boumah M Kambouris S A Mayouf K V Sheth W A Eid O Dowling J Harris M J Glucksman S Bahabri B F Meyer R J Desnick 《Nature genetics》2001,28(3):261-265
The inherited osteolyses or 'vanishing bone' syndromes are a group of rare disorders of unknown etiology characterized by destruction and resorption of affected bones. The multicentric osteolyses are notable for interphalangeal joint erosions that mimic severe juvenile rheumatoid arthritis (OMIMs 166300, 259600, 259610 and 277950). We recently described an autosomal recessive form of multicentric osteolysis with carpal and tarsal resorption, crippling arthritic changes, marked osteoporosis, palmar and plantar subcutaneous nodules and distinctive facies in a number of consanguineous Saudi Arabian families. We localized the disease gene to 16q12-21 by using members of these families for a genome-wide search for homozygous-by-descent microsatellite markers. Haplotype analysis narrowed the critical region to a 1.2-cM region that spans the gene encoding MMP-2 (gelatinase A, collagenase type IV; (ref. 3). We detected no MMP2 enzymatic activity in the serum or fibroblasts of affected family members. We identified two family-specific homoallelic MMP2 mutations: R101H and Y244X. The nonsense mutation effects a deletion of the substrate-binding and catalytic sites and the fibronectin type II-like and hemopexin/TIMP2 binding domains. Based on molecular modeling, the missense mutation disrupts hydrogen bond formation within the highly conserved prodomain adjacent to the catalytic zinc ion. 相似文献