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101.
Ryuji Fukuda Ryoji Yano Toshikazu Fukui Toshiharu Hase Akira Ishihama Hiroshi Matsubara 《Molecular & general genetics : MGG》1985,201(2):151-157
Summary In order to clone the Escherichia coli gene for the stringent starvation protein (SSP), we determined its N-terminal sequence as well as the sequence of two peptide fragments obtained by cyanogen bromide cleavage of the protein. We then chemically synthesized four sets of oligodeoxyribonucleotide mixtures that represented possible codon combinations for parts of these amino acid sequences. The synthetic oligonucleotides were labelled with 32P at their 5-termini and used as hybridization probes to detect DNA fragments containing the complementary sequences. Genomic Southern hybridization of E. coli chromosomal DNA gave up to ten DNA fragments hybridizing with each probe but only a few hybridized with two or more of the probes. The latter fragments were coloned in pBR322. By determining partial base sequences with a rapid method and examining proteins encoded by the DNA fragments, we were able to show that we had isolated a clone containing the complete SSP structural gene.Abbreviations SSP
stringent starvation protein
- PTH
phenylthiohydantoin 相似文献
102.
Nicole Houba-Hérin Hiroshi Hara Masayori Inouye Yukinori Hirota 《Molecular & general genetics : MGG》1985,201(3):499-504
Summary In order to determine the active site of penicillin-binding protein 3 of Escherichia coli (PBP3), the serine residue at position 307 was replaced with alanine, threonine or cysteine by oligonucleotide-directed site-specific mutagenesis. Since a unique BanII site exists at the position corresponding to serine-307, BanII digestion of the plasmid DNA after mutagenesis resulted in significant enrichment of the mutant plasmids. For mutagenesis, the gene coding for PBP3 (ftsI) was inserted into the expression cloning vector pIN-IIB. The hybrid protein produced was able to bind penicillin while mutant PBP3 in which serine-307 was replaced with either alanine or threonine did not lead to any detectable binding. However, contrary to the report of Broome-Smith et al. (1985) thiol-penicillin-binding protein 3, in which serine-307 was replaced with cysteine, was still able to bind penicillin. Replacement of serine-445 with an alanine residue had no effect on penicillin binding to PBP3. 相似文献
103.
The dependence of membrane potentials on changes in the extra-cellularK+ concentration [K+]e was investigated in potato tuber sliceswith dripping perfusion, and in growing Vigna hypocotyl segmentswith pressurized intra-organ perfusion methods. Only under anoxiawere the membrane potential of potato tuber slices and the electricpotential difference between the parenchyma symplast and xylem(Vpx) of Vigna hypocotyl segments depolarized markedly (46 mVand 42 mV/log[K+]e unit, respectively) with increasing [K+]eabove the critical values. The electric potential differencebetween the parenchyma symplast and organ surface (Vps of thehypocotyl segments remained nearly unchanged up to 30 mEq [K+]e.Under highly aerobic conditions the membrane potentials wererelatively independent of [K+]e except at very high K+ concentrations.Vps showed even hyperpolarization with the increasing KCl concentrationin the perfusion solution that is not in direct contact withthe surface membrane of the parenchyma symplast. The respiration-dependentelectrogenic components of the membrane potentials regularlyincreased with the increasing [K+]e. A voltage-dependent homeostaticcontrol of membrane potential is discussed. (Received August 13, 1984; Accepted December 21, 1984) 相似文献
104.
The heterogeneity of human fibroblasts as determined from the effects of hydrocortisone on cell growth and specific dexamethasone binding 总被引:1,自引:0,他引:1
To elucidate the heterogeneity of human fibroblasts from lung and skin, the effects of hydrocortisone on cell proliferation and the specific dexamethasone binding to cells were studied. Hydrocortisone at physiological concentrations stimulated the proliferation in three strains of human fetal lung fibroblasts and inhibited it in two strains. There are two kinds of fibroblasts in the human fetal lung in addition to the human skin fibroblasts reported previously. Dexamethasone-binding experiments showed that human fibroblasts may be classified into two groups with respect to the dissociation constant (Kd) of the binding reaction. The heterogeneity of human fibroblasts shown by Kd could not be correlated to classification on the basis of the effects of hydrocortisone on cell proliferation. The differences in Kd for the binding reactions suggest differences in donor tissues from which human fibroblasts are derived. 相似文献
105.
Yasunobu Antoku Tetsuo Sakai Kohsuke Tsukamoto Ikuo Goto Hiroshi Iwashita Yoshigoro Kuroiwa 《Journal of neurochemistry》1985,44(6):1667-1671
Phospholipid classes that included plasmalogens of erythrocyte membranes in seven myotonic dystrophy (MyD) patients and seven normal controls were analyzed by HPLC. No significant difference in phospholipid classes was found between patients with MyD and normal controls, but there was a visible difference in peak profiles of compounds of the phosphatidylethanolamine class. In the study of plasmalogens, we used two preparation methods: exposure to HCl and deacylation with mild alkaline. The area ratio of the plasmalogen form to the diacyl form in the phosphatidylethanolamine class of MyD erythrocyte membranes was significantly lower than that of normal controls. Fatty acid analyses showed that fatty acids of both phosphatidylethanolamine subclasses have less unsaturation in MyD. 相似文献
106.
The Escherichia coli cytochrome b556 gene, cybA, is assignable as sdhC in the succinate dehydrogenase gene cluster 总被引:2,自引:0,他引:2
Hiroshi Murakami Kiyoshi Kita Hiroshi Oya Yasuhiro Anraku 《FEMS microbiology letters》1985,30(3):307-311
Abstract The cytochrome b556 -deficient mutant Escherichia coli K12 strain TK3D11 [7] could not grow with succinate as the sole carbon source, but could grow well on dl -lactate. This finding suggested that cytochrome b556 is primarily responsible for oxidative metabolism and utilization of succinate. 24 Amino acid residues at the amino-terminal of purified cytochrome b556 were determined. This sequence coincided completely with amino acid residues 4 to 27, predicted from the DNA sequence of the sdhC gene, one of the unassigned open reading frames of the sdh gene cluster recently reported by Wood et al. [16]. Based on these and other results, we concluded that cybA , the gene for cytochrome b556 , is assignable as sdhC . 相似文献
107.
M. Tachibana H. Morioka M. Machino F. Mizukoshi O. Mizukoshi T. Yoshioka 《Histochemistry and cell biology》1985,82(2):197-199
Summary Triphosphoinositide (TPI), an aminoglycoside receptor and a possible regulator of cationic permeation through its ability to bind with Ca++, was localized by the protein-A gold technique in vestibular sensory epithelia using an antibody highly specific to TPI. TPI was detected on the stereocilia, kinocilia, and cuticular plate of hair cells, and in the reticular membrane of supporting cells. The cilia of hair cells are damaged by aminoglycosides at a relatively early stage of toxicity. Ca++-regulated bioactivity in this area is probably involved. 相似文献
108.
Evidence from morphology and anatomy (including embryology), as well as from palynology, chemistry and cytology, indicates
thatHydrastis is quite divergent from Ranunculaceae (in which the genus has been most often included) as well as from both Glaucidiaceae
and Berberidaceae. Distinctive features ofHydrastis, which demarcate it from Ranunculaceae but which are sometimes shared by Berberidaceae, are: the unique mode of origin of
the vascular supply to stamens and carpels; the micropyle being formed by both integuments; the xylem not V-shaped in cross
section; scalariform vessel perforations present; haploid chromosome number 13; pollen tectum consisting of a compound layer
of striae; leaf mesophyll not differentiated; the unique course of stem medullary bundles; D-galactose present. Its distinctive
higher haploid chromosome number, as well as its many less-specialized character states (in floral structure, leaf anatomy,
and xylem and vessel morphology), suggest thatHydrastis is a relictual primitive group which diverged early from a common ancestral stock of Ranunculaceae, Berberidaceae and probably
of Circaeasteraceae; at least some of the features shared byHydrastis and one or another of the families concerned seem to be a heritage from their common ancestor. We propose a reestablishment
of a monotypic family, Hydrastidaceae. 相似文献
109.
Spontaneous Mutations Modifying the Activity of Alcohol Dehydrogenase (Adh) in DROSOPHILA MELANOGASTER 总被引:2,自引:1,他引:1 下载免费PDF全文
In a marked-inversion balanced lethal system of the second chromosome of Drosophila melanogaster, mutations were accumulated under minimum pressure of natural selection in 1000 individual lines that originated essentially from two individuals. After about 300 generations, the specific activities of alcohol dehydrogenase of 69 randomly selected individual lines were measured with replications using four replicated vials (on 2 days—two replications per day) by observing the reduction of NAD+ to NADH at 340 nm. Total soluble protein as the basis of standardization of enzyme activity was measured by the Lowry method for each vial. A control experiment was made immediately after the establishment of 20 individual lines from a single genotype. A significant increase in genetic variance was observed among the mutation-accumulating lines but was not detected in the control experiment. The statistical analysis of the data on the basis of the one-band/one-gene hypothesis suggests that many mutations controlling the activity of alcohol dehydrogenase occurred in regions different from the alcohol dehydrogenase locus itself, mainly in the noncoding DNA. Furthermore, it is suggested that transposon-like elements are related to the induction of these changes in alcohol dehydrogenase specific activities. Additional experimental evidence supporting this conclusion is also given. 相似文献
110.
Yoshinobu Shioda Hiroshi Nagura Yutaka Tsutsumi Kazuo Shimamura Norikazu Tamaoki 《The Histochemical journal》1984,16(8):843-854
Summary Leu 7 immunoreactivity was demonstrated with the indirect peroxidase-labelled antibody method on frozen and paraffin-embedded tissue sections of human digestive organs. Anti-Leu 7 monoclonal antibody, which allegedly detects mononuclear cells with natural killer or killer activity, recognized lymphoid cells among intestinal epithelial cells and in the germinal centres of solitary lymphoid follicles of small and large intestine, and a few in gallbladder, liver and the lamina propria of the intestine. In addition, peripheral nerve fibres, endocrine cells in the gut and pancreas and carcinoid and islet cell tumours were also positively stained. At the ultrastructural level, Leu 7 antigen was localized on the plasma membrane of granulated lymphoid cells in the gut mucosa and on the secretory granules of intestinal endocrine cells. In normal pancreas, Leu 7 immunoreactivity was demonstrated in most cells containing pancreatic polypeptide and in many cells containing somatostatin or glicentin. Insulin-containing cells, however, lacked Leu 7 immunoreactant. These findings were obtained in both frozen sections and paraffin-embedded sections. The possible cross-reactivities of monoclonal antibodies are discussed as they raise an important caveat in immunohistochemical studies using these antibodies. 相似文献