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31.
32.
野生大豆基因文库的构建 总被引:4,自引:0,他引:4
以氯化铯密度梯度离心法纯化噬菌体λEMBL4,将纯化的EMBL4 DNA用BamH1/SalI双酶切制成载体。用CTAB(十六烷基三甲基溴化铵)法提取野生大豆(种名待定)大分子DNA,Sau3A部分酶解,从琼脂糖凝胶中回收10—22kb“目的”DNA片段,与载体连接,体外包装成重组噬菌体。所得重组子值为8×10(?)pfu(噬菌斑形成单位),达到了构建野生大豆基因文库要求的理论值。以栽培大豆7S贮藏蛋白a′-cDNA作探针,用噬菌斑原位杂交法从文库中筛选出一个阳性克隆。 相似文献
33.
Carlos Hermenegildo Goizane Marcaida Carmina Montoliu Santiago Grisolía María-Dolores Miñana Vicente Felipo 《Neurochemical research》1996,21(10):1237-1244
We proposed that acute ammonia toxicity is mediated by activation of NMDA receptors. To confirm this hypothesis we have tested
whether different NMDA receptor antagonists, acting on different sites of NMDA receptors, prevent death of mice induced by
injection of 14 mmol/Kg of ammonium acetate, a dose that induces death of 95% of mice. MK-801, phencyclidine and ketamine,
which block the ion channel of NMDA receptors, prevent death of at least 75% of mice. CPP, AP-5, CGS 19755, and CGP 40116,
competitive antagonists acting on the binding site for NMDA, also prevent death of at least 75% of mice. Butanol, ethanol
and methanol which block NMDA receptors, also prevent death of mice. There is an excellent correlation between the EC50 for preventing ammonia-induced death and the IC50 for inhibiting NMDA-induced currents. Acute ammonia toxicity is not prevented by antagonists of kainate/AMPA receptors, of
muscarinic or nicotinic acetylcholine receptors or of GABA receptors. Inhibitors of nitric oxide synthase afford partial protection
against ammonia toxicity while inhibitors of calcineurin, of glutamine synthetase or antioxidants did not prevent ammonia-induced
death of mice. These results strongly support the idea that acute ammonia toxicity is mediated by activation of NMDA receptors. 相似文献
34.
Summary To isolate a novel gene that contains an SH2 domain, we devised a rapid and nonradioactive cDNA library screening method using polymerase chain reaction (PCR). For PCR amplification, we designed degenerate oligonucleotide primers from the multialigned DNA sequences of SH2 domains. This method offers an inexpensive and efficient approach for the isolation of clones of interest from cDNA libraries. 相似文献
35.
E. A. Stewart A. White J. Tomfohrde S. Osborne-Lawrence L. Prestridge B. Bonne-Tamir I. H. Scheinberg P. St George-Hyslop M. Giagheddu J.-W. Kim J. K. Seo W. H.-y. Lo I. A. Ivanova-Smolenskaya S. A. Limborska L. L. Cavalli-Sforza L. A. Farrer A. M. Bowcock 《American journal of human genetics》1993,53(4):864-873
Wilson disease (WD), an autosomal recessive disorder of copper metabolism, has been previously mapped to chromosome 13q. Highly informative PCR-based polymorphic microsatellites closely linked to the WD locus (WND) at 13q14.3, as well as sequence-tagged sites for closely linked loci, are described. Two polymorphic microsatellite markers at D13S118 and D13S119 lie within 3 cM of WND. Two others (D13S227 and D13S228) were derived from a yeast artificial chromosome containing D13S31. These were placed on a genetic linkage map of chromosome 13 and were typed in 74 multiplex WD families from a variety of geographic origins (166 affected members). Multipoint analysis provides very high odds that the location of WND is between D13S31/D13S227/D13S228 and D13S59. Previous odds with RFLP-based markers were only 7:1 more likely than any other location. Current odds are 5,000:1. Preclinical testing of three cases of WD by using the highly informative polymorphic microsatellite markers is described. The markers described here ensure that 95% of predictive tests using DNA from both parents and from at least one affected sib will have an accuracy >99%. 相似文献
36.
Takahiro Okazaki Chiemi Nakanishi-Ito Naohiro Seo Takae Tanino Masafumi Takiguchi Kohji Egawa 《Cancer immunology, immunotherapy : CII》1993,36(2):83-88
Tumor-specific expression of Qa-2k antigen coded by the Q5k gene on various mouse tumor cells and immunological response of the host mice to the antigen have been demonstrated [Seo et al. (1992) J Exp Med 175: 547; Tanino et al. (1992) Cancer Immunol Immunother 35: 230]. The possibility was examined that Qa-2 antigen is one of the recognition target molecules of immunopotentiator-induced, H-2-nonrestricted tumoricidal lymphocytes of Qa-2– mice. Lymphocytes stimulated in vivo withP. acnes or culture-induced anomalous killers of B6.K1 mice did not exhibit significant in vitro cytotoxicity against B6.K1 lymphoblasts but lysed their Qa-2,3-congenic counterpart B6 lymphoblasts. To demonstrate the Qa-2 specificity of such cytotoxic cells more precisely, an L cell transformant clone (LQ7b/Kb), which expressed the 1 and 2 domains of the Qa-2 antigen (Q7b gene product), was generated by transfecting a cloned plasmid DNA containing a hybrid gene constructed from the 5 half of the Q7b gene and the 3 half of the H-2Kb gene (pQ7b/Kb). Using LQ7b/Kb cells as the target cells and the nylon-wool-nonadherent fraction of lymphocytes fromP. acnes-stimulated (C3H/He × B6.K1)F1 mice (H-2k, Qa-2–) as the effector cells of the in vitro cytotoxicity reaction, the presence of cytotoxic cells that recognize the 1/2 region of the Q7b gene product was demonstrated. The cytotoxic activity was dependent on T cells bearing T cell receptors of the / type (TCR/). The (C3H/He × B6.K1)F1 effector cells, as well as the B6.K1 effector cells also lysed BW5147 lymphoma cells (Qa-2k+) derived from AKR mice (Qa-2–, H-2k). By target-competition experiments it was shown that some of the effector cells lytic to BW5147 were identical to those that lysed LQ7b/Kb. Therefore some of the tumoricidal cells induced by the immunopotentiator interact with the target tumor cells through recognition of the 1/2 region of the Qa-2k tumor antigen by TCR/. 相似文献
37.
Hybrid cells were obtained from somatic cell fusion among male, female, and tetrasporangial plants in Griffithsia japonica Okamura by a wound-healing process. Isolated fusion cells regenerated new mature plants with mixed reproductive structures. The plants regenerated from hybrid cells between male and female plants developed into 1) spermatangiate, 2) carpogonial, 3) bisexual with spermatangia and carpogonial branches, 4) mixed-phase with spermatangia and tetrasporangia, or 5) bisexual/mixed-phase plants with spermatangia, carpogonial branches, and tetrasporangia. About 70% of the plants regenerated from hybrid cells between male and female plants produced tetrasporangia that were always formed with spermatangia on a single cell. Some of those tetrasporangia released tetraspores, six of which gave rise to mature plants. The plants regenerated from hybrid cells between male and tetrasporangial plants developed into spermatangiate, tetrasporangiate, or mixed-phase plants with spermatangia and tetrasporangia. The plants regenerated from hybrid cells between female and tetrasporangial plants developed into carpogonial, tetrasporangiate, or mixed-phase plants with carpogonial branches and tetrasporangia. All types of reproductive structures we re functional. 相似文献
38.
Summary A new process for the production of small size dextran is developed in which dextran is produced by cultures of Leuconostoc mesenteroides in the presence of a partially constitutive mutant of Lipomyces starkeyi producing dextranase. Mixed cultures were examined by scanning electron microscopy with ruthenium to show the effects of the mixed culture on low molecular weight dextran (M.W. of 5,000 – 100,000) formation. The presence of the size variation in dextran was confirmed by gel permeation chromatography. 相似文献
39.
本文用聚丙烯酰胺凝胶电泳方法,对瑞氏七鳃鳗五种不同组织(骨骼肌、心、肾、肠、鳃)中LDH同工酶进行了分析研究,结果表明,LDH同工酶具有组织特异性,其中骨骼肌中含有五种LDH同。酶,即LDH1、LDH2、LDH3、LDH4、LDH5,鳃含有LDH1和LDH4而肾和心只含有LDH1,肠只含有LDH4。 相似文献
40.
HhaI and HpaII DNA methyltransferases bind DNA mismatches, methylate uracil and block DNA repair. 总被引:10,自引:7,他引:3
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The hydrolytic deamination of 5-methylcytosine (5-mC) to thymine (T) is believed to be responsible for the high mutability of the CpG dinucleotide in DNA. We have shown a possible alternate mechanism for mutagenesis at CpG in which HpaII DNA-(cytosine-5) methyltransferase (M.HpaII) can enzymatically deaminate cytosine (C) to uracil (U) in DNA [Shen, J.-C., Rideout, W.M., III and Jones, P.A., Cell, 71, 1073-1080, (1992)]. Both the hydrolytic deamination of 5-mC and enzymatic deamination of C create premutagenic DNA mismatches (G:U and G:T) with the guanine (G) originally paired to the normal C. Surprisingly, we found that DNA-(cytosine-5) methyltransferases have higher affinities for these DNA mismatches than for their normal G:C targets and are capable of transferring a methyl group to the 5-position of U, creating T at low efficiencies. This binding by methyltransferase to mismatches at the recognition site prevented repair of G:U mismatches by uracil DNA glycosylase in vitro. 相似文献