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951.
Selective inhibition of splicing at the avian sarcoma virus src 3' splice site by direct-repeat posttranscriptional cis elements 下载免费PDF全文
The direct-repeat elements (dr1) of avian sarcoma virus (ASV) and leukosis virus have the properties of constitutive transport elements (CTEs), which facilitate cytoplasmic accumulation of unspliced RNA. It is thought that these elements represent binding sites for cellular factors. Previous studies have indicated that in the context of the avian sarcoma virus genome, precise deletion of both ASV dr1 elements results in a very low level of virus replication. This is characterized by a decreased cytoplasmic accumulation of unspliced RNA and a selective increase in spliced src mRNA. Deletion of either the upstream or downstream dr1 results in a delayed-replication phenotype. To determine if the same regions of the dr1 mediate inhibition of src splicing and unspliced RNA transport, point mutations in the upstream and downstream elements were studied. In the context of viral genomes with single dr1 elements, the effects of the mutations on virus replication and increases in src splicing closely paralleled the effects of the mutations on CTE activity. For mutants strongly affecting CTE activity and splicing, unspliced RNA but not spliced RNA turned over in the nucleus more rapidly than wild-type RNA. In the context of wild-type virus containing two dr1 elements, mutations of either element that strongly affect CTE activity caused a marked delay of virus replication and a selective increase in src splicing. However, the turnover of the mutant unspliced RNA as well as the spliced mRNA species did not differ significantly from that of the wild type. These results suggest the dr1 elements in ASV act to selectively inhibit src splicing and that both elements contribute to the fitness of the wild-type virus. However, a single dr1 element is sufficient to stabilize unspliced RNA. 相似文献
952.
A single pair of oligonucleatide primer selected within a highly conserved region of the DNA polymerase gene in herpesviruses was synthesized. The competitive template DNA purified from cytomegalovirus (CMV) DNA was used to carry out competiitve PCR amplification with herpes simplex virus type 1 (HSV1) DNA (target sequences). And anti-HSV1 effects of acyclovir (ACV) was investigated by the method.The results showed that the efficacy of PCR amplification was equal to each other(the ratio of the quantity of c… 相似文献
953.
954.
Auxin Deprivation Induces Synchronous Golgi Differentiation in
Suspension-Cultured Tobacco BY-2 Cells 总被引:7,自引:1,他引:6 下载免费PDF全文
To date, the lack of a method for inducing plant cells and their Golgi stacks to differentiate in a synchronous manner has made it difficult to characterize the nature and extent of Golgi retailoring in biochemical terms. Here we report that auxin deprivation can be used to induce a uniform population of suspension-cultured tobacco (Nicotiana tabacum cv BY-2) cells to differentiate synchronously during a 4-d period. Upon removal of auxin, the cells stop dividing, undergo elongation, and differentiate in a manner that mimics the formation of slime-secreting epidermal and peripheral root-cap cells. The morphological changes to the Golgi apparatus include a proportional increase in the number of trans-Golgi cisternae, a switch to larger-sized secretory vesicles that bud from the trans-Golgi cisternae, and an increase in osmium staining of the secretory products. Biochemical alterations include an increase in large, fucosylated, mucin-type glycoproteins, changes in the types of secreted arabinogalactan proteins, and an increase in the amounts and types of molecules containing the peripheral root-cap-cell-specific epitope JIM 13. Taken together, these findings support the hypothesis that auxin deprivation can be used to induce tobacco BY-2 cells to differentiate synchronously into mucilage-secreting cells. 相似文献
955.
Plant regeneration through organogenesis from callus induced from mature zygotic embryos of loblolly pine 总被引:8,自引:1,他引:7
Mature zygotic embryos from three seed sources of loblolly pine were cultured on callus induction medium containing 10 mg
l–1
α-naphthaleneacetic acid, 4 mg l–1 benzyladenine (BA), 400 mg l–1 casein hydrolysate, and 400 mg l–1 glutamine for 6 weeks. Light-yellow, loose, glossy, globular callus was formed, and the highest frequency was 35.7%. The
highest differentiation frequency of callus on adventitious bud induction medium was 62.1%. After culture of calli with adventitious
buds on elongation medium for 6 weeks, adventitious shoots more than 1.0 cm in height were selected for rooting. On rooting
medium supplemented with 0.1 mg l–1 indole-3-butyric acid, 1 mg l–1 BA, and 0.5 mg l–1 gibberellic acid, the highest rooting frequency of adventitious shoots was 46% in a culture period of 6 weeks. Established
plants survived following transfer to soil at a frequency of 71%.
Received: 14 May 1997 / Revision received: 25 September 1997 / Accepted: 11 October 1997 相似文献
956.
火炬松胚性细胞悬浮培养物的生长参数变化 总被引:1,自引:0,他引:1
以火炬松(PinustaedaL.)成熟合子胚来源的胚性愈伤组织为材料建立了胚性细胞悬浮系,测定了其培养物的鲜重、干重、细胞体积和胚数及培养液中的pH值、电导率和蔗糖浓度等生长参数在培养过程中的变化动态。结果表明,在培养周期内,培养液中的pH值、电导率和蔗糖浓度的逐步降低与培养物的鲜重、干重、细胞体积和胚胎数的逐步增加保持一致性。在培养至18—21d,pH值、电导率和蔗糖浓度均接近或降到最低点,而胚数及细胞体积的增长都达到最高点。 相似文献
957.
黑曲霉T21是由黑曲霉3.795经诱变育种获得的糖化酶高产菌株,为阐明其高产的分子机制,由黑曲霉3.795克隆了糖化酶结构基因及其5′旁侧序列,并与黑曲霉T21的相应序列进行了比较.由黑曲霉3.795菌丝体分离染色体DNA,Southern杂交分析表明,糖化酶结构基因位于~2.5kb的EcoRⅠ-EcoRⅤ染色体DNA片段上,在此EcoRⅠ位点上游约1.0kb处有一SalⅠ位点.为构建糖化酶结构基因及其5′旁侧序列的基因组文库,该染色体DNA分别用EcoRⅠ+EcoRⅤ和EcoR+SalⅠ消化,琼脂糖凝胶电泳分离并回收长度在1.0kb左右和2.5kb左右的DNA片段,分别与pUC19载体连接后转化入E.coliDH5.用原位杂交方法筛选到了携带糖化酶基因编码区及其1505bp5′旁侧序列的阳性克隆.对克隆片段的DNA序列进行了测定并与黑曲霉T21的相应序列进行了比较,结果表明,在糖化酶基因编码区及其150bp3′非编码区内,未发现碱基差异,但在-340~-1505的5′上游区内发生了9个位置的碱基变化,包括缺失、插入和替换.这些结果表明,黑曲霉T21与3.795的糖化酶产量的差异与其结构基因无关,但可能与其 相似文献
958.
959.
Dominant Negative Alleles of SEC10 Reveal Distinct
Domains Involved in Secretion and Morphogenesis in Yeast 总被引:2,自引:1,他引:1 下载免费PDF全文
The accurate targeting of secretory vesicles to distinct sites on the plasma membrane is necessary to achieve polarized growth and to establish specialized domains at the surface of eukaryotic cells. Members of a protein complex required for exocytosis, the exocyst, have been localized to regions of active secretion in the budding yeast Saccharomyces cerevisiae where they may function to specify sites on the plasma membrane for vesicle docking and fusion. In this study we have addressed the function of one member of the exocyst complex, Sec10p. We have identified two functional domains of Sec10p that act in a dominant-negative manner to inhibit cell growth upon overexpression. Phenotypic and biochemical analysis of the dominant-negative mutants points to a bifunctional role for Sec10p. One domain, consisting of the amino-terminal two-thirds of Sec10p directly interacts with Sec15p, another exocyst component. Overexpression of this domain displaces the full-length Sec10 from the exocyst complex, resulting in a block in exocytosis and an accumulation of secretory vesicles. The carboxy-terminal domain of Sec10p does not interact with other members of the exocyst complex and expression of this domain does not cause a secretory defect. Rather, this mutant results in the formation of elongated cells, suggesting that the second domain of Sec10p is required for morphogenesis, perhaps regulating the reorientation of the secretory pathway from the tip of the emerging daughter cell toward the mother–daughter connection during cell cycle progression. 相似文献
960.
The effects of microhabitat differentiation on small-scale plant community structure in the Chiuhuahuan Desert were studied using multivariate analysis. The results showed that microhabitats (i.e., kangaroo rat mounds, ant mounds, shrubs, half-shrubs, and open areas) played a critical role in structuring small-scale plant community structure and maintaining species diversity. Annual plants were much more sensitvive to the presence of differentiated microhabitats than perennials and winter annuals exhibited stronger microhabitat perferences than summer annuals. Species diversity was highest on ant mounds while open areas supported the lowest diversity during both winter and summer. Biomass was highest in the shrub habitats followed by kangaroo rat mounds, ant mounds, half-shrubs, and open areas. Much of the diversity of these plants could be explained by the individualistic responses of species to the biotic effect of other plants or to disturbance by animals, or individualistic responses of species to differences in microenvironments. 相似文献