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921.
Photo-CIDNP study of the interaction between the glucocorticoid receptor DNA-binding domain and glucocorticoid response elements 总被引:1,自引:0,他引:1
E. Kellenbach T. Härd R. Boelens K. Dahlman J. Carlstedt-Duke J. -Å. Gustafsson G. A. van der Marel J. H. van Boom B. Maler K. R. Yamamoto R. Kaptein 《Journal of biomolecular NMR》1991,1(1):105-110
Summary Photo-CIDNP studies were performed on two protein fragments that both contain the double zinc-finger DNA-binding domain of the glucocorticoid receptor. In the absence of DNA, Tyr452 and Tyr474 are polarised in both fragments while Tyr497 is not. Addition of a palindromic glucocorticoid response element (GRE) results in the suppression of Tyr474 polarization while the polarization of Tyr452 is unaffected. The same result is observed upon adding a half GRE to the protein fragment indicating that the suppression of Tyr474 polarization is not due to protein-protein contacts but to interaction with DNA. 相似文献
922.
923.
Mechanisms of somatic embryogenesis in cell cultures: Physiology,biochemistry, and molecular biology
A. Komamine R. Kawahara M. Matsumoto S. Sunabori T. Toya A. Fujiwara M. Tsukahara J. Smith M. Ito H. Fukuda K. Nomura T. Fujimura 《In vitro cellular & developmental biology. Plant》1992,28(1):11-14
Summary One of the most characteristic cell functions in plants is totipotency. Somatic embryogenesis can be regarded as a model system
for the investigation of mechanisms of totipotency, because a high frequency and synchronous embryogenic system from single
somatic cells has been established in carrot suspension cultures. Four phases are recognized in this process, and several
molecular markers, viz. polypeptides, mRNAs, antigens against monoclonal antibodies, can be detected during the expression
of totipotency, but they disappear during its loss. Four organ-specific genes have been isolated from hypocotyls and roots
by differential screening. They were expressed preferentially after the globular-heart stages of embryogenesis, and were strongly
suppressed by auxin. A CEM 1 gene was isolated by differential screening of embryogenic cell clusters. This gene was expressed
strongly and transiently during the proglobular and globular stages. The sequence of CEM 1 was found to encode a polypeptide
showing high homology to the elongation factor isolated from eucaryotic cells. Thus good progress is being made in understanding
the basic mechanisms of somatic embryogenesis.
Presented in the Session-in-Depth Developmental Biology of Embryogenesis at the 1991 World Congress on Cell and Tissue Culture,
Anaheim, California, June 16–20, 1991. 相似文献
924.
Elsayed A. M. Abdallah David A. Jett Mohyee E. Eldefrawi Amira T. Eldefrawi 《Journal of biochemical and molecular toxicology》1992,7(2):125-132
The effects of the organophosphorus anticholinesterase paraoxon on the binding of radioactive ligands to the M3 subtype of the muscarinic receptor and receptor-coupled synthesis of second messengers in intact rat submaxillary gland (SMG) cells were investigated. The binding of [3H]quinuclidinyl benzilate ([3H]QNB) was most sensitive to atropine and the M3-specific antagonist 4-DAMP followed by pirenzepine and least sensitive to the cardioselective M2 antagonist AFDX116. This, and the binding characteristics of [3H]4-DAMP, confirmed that the muscarinic receptors in this preparation are of the M3 subtype. Activation of these muscarinic receptors by carbamylcholine (CBC) produced both stimulation of phosphoinositide (PI) hydrolysis and inhibition of cAMP synthesis, suggesting that this receptor subtype couples to both effector systems. Paraoxon (100 μM) reduced Bmax of [3H]4-DAMP binding from 27 ± 4 to 13 ± 3 fmol/mg protein with nonsignificant change in affinity, suggesting noncompetitive inhibition of binding by paraoxon. Like the agonist CBC, paraoxon inhibited the forskolininduced cAMP formation in SMG cells with an EC50 of 200 nM, but paraoxon was > 500 fold more potent than CBC. However, while the inhibition by CBC was counteracted by 2 μM atropine, that by paraoxon was unaffected by up to 100 μM atropine. It suggested that this effect of paraoxon was not via binding to the muscarinic receptor. Paraoxon did not affect β-adrenoreceptor function in the preparation, since it did not affect the 10 μM isoproterenol-induced cAMP synthesis, which was inhibited totally by 10 μM propranolol and partially by CBC. Paraoxon had a small but significant effect on CBC-stimulated PI metabolism in the SMG cells. It is suggested that paraoxon binds to two different sites in these SMG cells. One is an allosteric site on the M3 muscarinic receptor which affects ligand binding and may modulate receptor function. The other site may be on the Gi proteinadenylyl cyclase system, and produces CBC-like action, that is, inhibition of the forskolin-stimulated [3H]cAMP synthesis, and is unaffected by atropine inhibition of the muscarinic receptor. This adds to the complexity of paraoxon actions on muscarinic receptors and their effector systems. 相似文献
925.
Pierangelo Luporini Cristina Miceli Claudio Ortenzi Adriana Vallesi 《Genesis (New York, N.Y. : 2000)》1992,13(1):9-15
Euplotes raikovi, like other ciliates, passes through a postconjugal immaturity, operatively identified by an apparent cell inability to form mating pairs under experimental conditions that are the same as those used for inducing mating at maturity. In cells homozygous for the gene mat-2, which controls the pheromone Er-2, Er-2 mRNA synthesis and mature Er-2 secretion were shown to start from the very beginning of the life cycle and continue throughout immaturity, although to extents estimated to be 5- to 10-fold lower than at maturity. In addition, experiments of 125 I-Er-2 binding and crosslinking provided evidence that autocrine pheromone-binding sites, showing values of the dissociation constant of the order of 10?9 M, are on the surface of immature cells. The number of these sites per cell was estimated to increase from less than 106 per cell of 5–7 fissions of age, to about 16 × 106 at maturity. These results were taken to suggest that a pheromone-receptor production is stimulated during immaturity by autocrine pheromone binding to cells and that this production might be essential for the development of a pheromone-receptor density high enough to transform the cell from “immature” to “adult,” that is competent to respond as well to pheromones of conspecific, genetically different cells. © 1992 Wiley-Liss, Inc. 相似文献
926.
本文是26篇关于丝状真菌基因表达系统的研究论文的综述,包括两部份内容。前一部分叙述1979年开始建立并迅速发展起来的丝状真菌转化系统,着重介绍丝状真菌中转化系统的构建及转化的一般特点。后一部分叙述在转化系统发展基础上产生的丝状真菌基因工程,文中列出了截至1991年9月为止报道的一些成功的实例,说明它在丝状真菌工业育种和作为外源基因产物的生产和分泌系统中的应用。 相似文献
927.
Low doses of estradiol, administered as pulses, are as effective as higher doses for priming ovariectomized (OVX) guinea pigs to display progesterone-facilitated lordosis. High doses of estradiol, administered by constant-release implants, induce progestin receptors in many substance P-immunoreactive (SP-IR) neurons in the ventrolateral hypothalamus (VLH), a site at which estradiol primes OVX guinea pigs to respond behaviorally to progesterone. To test the hypothesis that behaviorally effective estradiol pulses induce progestin receptors selectively in substance P-containing neurons in the VLH, OVX females received estradiol implants 1 week prior to perfusion, or two pulses of estradiol- 17β, injected 39 and 11 h before perfusion. Colchicine was administered intracerebroventricularly prior to perfusion. No significant differences were observed in the total number of progestin receptor-immunoreactive (PR-IR) or substance P-immunoreactive cells in the VLH and VLH/ventromedial hypothalamus (VMH), respectively, of females receiving the two estradiol treatments. However, the percentage of PR-IR cells in the VLH also immunoreactive for SP was significantly higher in the estradiol pulse-treated (53%), than in the estradiol capsule-implanted animals (36%). These data suggest that behaviorally effective estradiol pulses induce progestin receptors selectively in substance P-containing neurons in the VLH and are consistent with the hypothesis that substance P is involved in progesterone-facilitated lordosis in guinea pigs. 相似文献
928.
J P van Leeuwen J C Birkenh?ger C J Buurman J P Schilte H A Pols 《FEBS letters》1990,270(1-2):165-167
In several cell types 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) causes up-regulation of its receptor. The present study demonstrates that in the osteoblast-like cell line UMR 106 this up-regulation is inhibited by two different calcium channel blockers (nitrendipine, verapamil). Also with chelating extracellular calcium (EGTA) and by inhibition of calcium release from intracellular stores (TMB-8) comparable results were obtained. These findings indicate that calcium is functionally involved in this cellular response to the steroid hormone 1,25(OH)2D3. Moreover, data obtained with EGTA show that the 1,25(OH)2D3 receptor level is closely regulated by the extracellular calcium concentration. 相似文献
929.
A simple method for shortening a plasmid genome using a system of plasmid cointegration mediated by a Tn3 mutant 总被引:3,自引:0,他引:3
This paper describes a simple method for the isolation of small plasmids of various sizes from pSMI, a derivative of the resistance plasmid R 100. The method is based on the observation that a repressor-negative mutant of the ampicillin-resistance (ampr) transposon Tn3, Tn3 No. 5, mediates cointegration of a plasmid carrying Tn 3 No. 5 (pMB8::Tn 3 No. 5) into virtually any site on pSMI. The resulting cointegrate plasmids contain the pSMI sequence which is joined with the ampr gene of the Tn 3 mutant. This cointegration is so frequent that large cointegrate plasmids can be readily detected in the total plasmid DNA prepared from cells carrying pSMI and pMB8::Tn3 No. 5. We were able to isolate small plasmids of various sizes by digesting the total plasmid DNAs with restriction endonucleases which cut both pSM 1 and Tn3 No. 5 sequences present in the cointegrates and subsequently ligating the restriction fragment containing both the ampr gene and the region necessary for replication of pSMI. Analysis of these plasmids, named pBV plasmids, with restriction endonucleases and by nucleotide sequencing allowed us to determine regions necessary or unnecessary for replication, thus defining a minimal replication region of pSMI. The present method is generally useful for the isolation of small derivatives from any large plasmid for the study of genes and sites adjacent to or within the minimal replication region of the plasmid. 相似文献
930.