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31.
Susana Valenciano J. Ramón De Lucas A. Pedregosa Inmaculada F. Monistrol F. Laborda 《Archives of microbiology》1996,166(5):336-341
Aspergillus nidulans is able to grow on oleic acid as sole carbon source. Characterization of the oleate-induced β-oxidation pathway showed the
presence of the two enzyme activities involved in the first step of this catabolic system: acyl-CoA oxidase and acyl-CoA dehydrogenase.
After isopicnic centrifugation in a linear sucrose gradient, microbodies (peroxisomes) housing the β-oxidation enzymes, isocitrate
lyase and catalase were clearly resolved from the mitochondrial fraction, which contained fumarase. Growth on oleic acid was
associated with the development of many microbodies that were scattered throughout the cytoplasm of the cells. These microbodies
(peroxisomes) were round to elongated, made up 6% of the cytoplasmic volume, and were characterized by the presence of catalase.
The β-oxidation pathway was also induced in acetate-grown cells, although at lower levels; these cells lacked acyl-CoA oxidase
activity. Nevertheless, growth on acetate did not cause a massive proliferation of microbodies in A. nidulans.
Received: 8 March 1996 / Accepted: 5 August 1996 相似文献
32.
J. Ramón De Lucas Susana Valenciano A. I. Domínguez Geoffrey Turner F. Laborda 《Archives of microbiology》1997,168(6):504-512
Conidia of Aspergillus nidulans were mutagenized with ultraviolet light and were incubated on a special selective medium containing the catalase inhibitor
3-amino-1,2,4-triazole. From approximately 5 × 107 viable UV-irradiated conidia tested, 423 stable mutants resistant to 3-amino-1,2,4-triazole were recovered, of which 40
were unable to grow on minimal medium with oleic acid as the sole carbon source. These oleate-nonutilizing (Ole–) mutants did not grow on medium with carbon sources requiring functional peroxisomes (oleate, butyrate, acetate, or ethanol),
but grew well on medium with carbon sources supposedly not requiring such organelles (glucose, glycerol, l-glutamate, or l-proline). The Ole– mutants carried mutations in one of five nuclear genes affecting acetate utilization: acuJ, acuH, acuE, acuL, and perA. The perA21 strain (DL21) carried a mutation in a gene that is not allelic with any of the known acu loci and displayed a phenotype resembling that described in the Pim– (peroxisome import defective) mutants of Hansenula polymorpha. Hyphae of the perA21 mutant contained a few small peroxisomes with the bulk of peroxisomal enzymes remaining in the 20,000 ×g supernatant, but produced wild-type levels of penicillin.
Received: 16 April 1997 / Accepted: 26 July 1997 相似文献
33.
N-Linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells 总被引:5,自引:3,他引:2
The potential of insect cell cultures and larvae infected with recombinant
baculoviruses to produce authentic recombinant glycoproteins cloned from
mammalian sources was investigated. A comparison was made of the N-linked
glycans attached to secreted alkaline phosphatase (SEAP) produced in four
species of insect larvae and their derived cell lines plus one additional
insect cell line and larvae of one additional species. These data survey
N-linked oligosaccharides produced in four families and six genera of the
order Lepidoptera. Recombinant SEAP expressed by recombinant isolates of
Autographa californica and Bombyx mori nucleopolyhedroviruses was purified
from cell culture medium, larval hemolymph or larval homogenates by
phosphate affinity chromatography. The N-linked oligosaccharides were
released with PNGase-F, labeled with 8- aminonaphthalene-1-3-6-trisulfonic
acid, fractionated by polyacrylamide gel electrophoresis, and analyzed by
fluorescence imaging. The oligosaccharide structures were confirmed with
exoglycosidase digestions. Recombinant SEAP produced in cell lines of
Lymantria dispar (IPLB-LdEIta), Heliothis virescens (IPLB-HvT1), and Bombyx
mori (BmN) and larvae of Spodoptera frugiperda, Trichoplusia ni ,
H.virescens , B.mori , and Danaus plexippus contained oligosaccharides that
were structurally identical to the 10 oligosaccharides attached to SEAP
produced in T.ni cell lines. The oligosaccharide structures were all
mannose-terminated. Structures containing two or three mannose residues,
with and without core fucosylation, constituted more than 75% of the
oligosaccharides from the cell culture and larval samples.
相似文献
34.
Wall formation inSaccharomyces cerevisiae seems to be the result of two main patterns of wall material deposition: (i) around the whole periphery of the cell in nonbudding
ones, and (ii) mainly at the tip of the daughter cell or at the cross wall that separates dividing cells. This interpretation
has been obtained following experiments in which RNA or protein synthesis has been inhibited. Under these conditions, glucan
formation takes place, and wall thickening is probably due to the accumulation of this polysaccharide. Furthermore, once a
pattern of wall deposition has been established, it is not modified by inhibition of RNA or protein synthesis. 相似文献
35.
In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold 总被引:14,自引:4,他引:10 下载免费PDF全文
In situ hybridization has become a standard method for localizing DNA or RNA sequences in cytological preparations. We developed two methods to extend this technique to the transmission electron microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope in situ hybridization. Radioactively labeled complementary RNA (cRNA) is hybridized to metaphase chromosomes deposited on electron microscope grids and fixed in 70 percent ethanol vapor; hybridixation site are detected by autoradiography. Specific and intense labeling of chromosomal centromeric regions is observed even after relatively short exposure times. Inerphase nuclei present in some of the metaphase chromosome preparations also show defined paatterms of satellite DNA labeling which suggests that satellite-containing regions are associate with each other during interphase. The sensitivity of this method is estimated to at least as good as that at the light microscope level while the resolution is improved at least threefold. The second method, which circumvents the use of autoradiogrphic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction is improved at least threefold. The second method, which circumvents the use of autoradiographic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction with an antibody against biotin and secondary antibody adsorbed to the surface of over centromeric heterochromatin and along the associated peripheral fibers. Labeling is on average ten times that of background binding. This method is rapid and possesses the potential to allow precise ultrastructual localization of DNA sequences in chromosomes and chromatin. 相似文献
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39.
Antonio Ramos Martínez Beatriz Orden Martínez Jessica Polo Laborda Blanca García Magallón Mónica Fernández Castro Carlos Ortega Sánchez Manuel Gil Navarro 《Revista iberoamericana de micología》2012,29(4):241-244
BackgroundFungal arthritis is usually of haematogenous origin, and mainly affects patients with impaired cellular immunity or users of intravenous drugs. The infection in immunocompetent patients is generally caused by direct inoculation of the microorganism through an invasive device. The experience of azole therapy in these patients is limited.Case reportWe report a case of arthritis caused by Scedosporium apiospermum characterized by its slow onset, lack of response to posaconazole and caspofungin, and its successful resolution after surgical debridement and treatment with voriconazole.ConclusionsTreatment with voriconazole and surgical debridement is an effective therapy for arthritis due to S. apiospermum. 相似文献
40.
dlk1 is an epidermal growth factor (EGF)-like homeotic protein containing an intracellular region, a single transmembrane domain, and an extracellular region possessing six EGF-like repeats and a protease-target sequence. dlk1 functions as a modulator of adipogenesis, and other differentiation processes. The molecular mechanisms by which dlk1 regulates these processes are unclear. It has been reported that different Dlk1 mRNA spliced variants, encoding for isoforms possessing the protease-target sequence or not, determine the production of membrane-associated or soluble, secreted extracellular dlk1 proteins that appear to affect adipogenesis of 3T3-L1 cells differently. In particular, only soluble variants inhibit this process. Some recent evidence suggest that dlk1 may modulate extracellular stimuli inducing differentiation. Thus, an enforced decrease of Dlk1 expression in BALB/c 3T3 cells, which results in an increase of their adipogenic potential in response to insulin-like growth factor 1 (IGF-1), modifies the kinetics and levels of activation of ERK1/2 triggered by it. In this work, we identified a strong and specific interaction between the protease-target dlk1 region and the non-IGF binding region of IGF binding protein 1 (IGFBP1), a protein that binds to IGFs and modulates their action. We also observed that the increased adipogenic potential of 3T3-L1 cells caused by diminishing Dlk1 expression through transfection with an antisense Dlk1 expression construct was inhibited by the presence of IGFBP1 in the differentiation medium. On the other hand, the presence of IGFBP1 in the culture medium slightly increased the adipogenic potential of control 3T3-L1 cells, expressing regular levels of Dlk1. These data suggest that membrane dlk1 variants bind to extracellular IGFBP1/IGF-1 complexes, which may favor the release of IGF-1 and increase the local concentration of free IGF-1 that can enhance IGF receptor signaling, leading to adipogenesis. 相似文献