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241.
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Heterologous expression of an engineered truncated form of human Lewis fucosyltransferase (Fuc-TIII) by the methylotrophic yeast Pichia pastoris 总被引:1,自引:0,他引:1
Gallet PF; Vaujour H; Petit JM; Maftah A; Oulmouden A; Oriol R; Le Narvor C; Guilloton M; Julien R 《Glycobiology》1998,8(9):919-925
A stable GS115 Pichia pastoris recombinant strain was constructed to
secrete a truncated form of the human alpha(1,3/4) fucosyltransferase
(amino acids 45-361). Enzyme production resulted from a secretory pathway
based on the pre-pro- alpha mating factor signal sequence of the yeast
Saccharomyces cerevisiae . Following its transit through the Golgi
apparatus, the enzyme accumulated in the periplasmic space before its
release in the culture broth (about 30 mg/l). Cell-enclosed enzyme (
approximately 0.16%) proved to be fairly stable for many freezing and
thawing cycles and could be used several times as an immobilized catalyst.
Soluble enzyme (>99.8%) representing the main protein of the culture
broth (10%) has been characterized by Western-blotting, substrate
specificities and kinetic parameters. The two forms (cell- enclosed and
soluble) of recombinant enzyme may be used for in vitro synthesis of
Lewisadeterminants.
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243.
Size homoplasy and mutational processes of interrupted microsatellites in two bee species, Apis mellifera and Bombus terrestris (Apidae) 总被引:6,自引:0,他引:6
Similar microsatellite electromorphs (PCR products of the same size) can
arise from independent mutational events. Such alleles are not identical by
descent. This phenomenon, termed size homoplasy, was studied by sequencing
electromorphs of two microsatellite loci in which the stretch of basic
repeats is interrupted by different short (1-2 bp) DNA motifs. The number
and position of these interruptions were established for electromorphs from
closely and distantly related populations of honeybees and bumblebees. No
sequence difference was found when electromorphs came from the same
subspecies or from closely related subspecies, suggesting that they were
probably identical by descent. In contrast, sequence differences were often
detected in distantly related subspecies, showing that size homoplasy
frequently occurs at this level of population differentiation. Size
homoplasy is increased by limits to free length variation of alleles, a
phenomenon that seems to act on interrupted microsatellites when comparing
distantly related taxa, that is, honeybee subspecies from different
evolutionary lineages. Electromorph sequences suggest that, within the
scope of these limits, large mutation events have occurred frequently at
both interrupted loci studied. In good agreement with the molecular data,
computations based on the observed heterozygosity and number of
electromorphs and simulation studies showed that neither locus fits the
one-step stepwise mutant model (SMM). We speculate that interrupted
microsatellites in general could be characterized by a higher variance in
repeat number and consequently a lower homoplasy rate than pure ones.
Hence, interrupted microsatellites should be most appropriate for
investigating population differentiation and evolutionary relationship
between relatively distant populations.
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Although invertebrate drift is an important ecological process in lotic ecosystems, very little is known about it in Kenyan rivers. The primary aim of this study was to investigate the effect of driftnet mesh size and exposure duration on drift density in 2017. Drift samples were dominated by Chironomidae, Baetidae, Simuliidae, Caenidae and Culicidae. The 100 µm mesh driftnet had the highest mean invertebrate density, followed by the 250 µm and 500 µm nets. Invertebrate drift densities decreased with increased exposure time. This study demonstrates that sampler mesh size and exposure time should be taken into account when characterising invertebrate drift in streams. Future studies should consider sampling different biotopes and during different seasons. 相似文献