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从江浙蝮蛇毒腺中抽提总RNA,RT-PCR进行体外扩增,获得江浙蝮蛇蛇毒蛋白C激活因子基因,克隆至pGEX-5X-3载体中,对3个重组克隆分别作DNA全序列分析,通过遗传密码推导出相应的氨基酸序列,与其它已知的丝氨酸复白酶蛇毒蛋白的氨基酸作比较,其中许多上氨基酸有很强的同源性,该基因的成功克隆,不仅推导出江浙蝮蛇蛇毒蛋白C激活因子的蛋白质序列,也为进一步开展江浙蝮蛇蛇毒蛋白C激活因子蛋白质工程的研究工作打下良好的基础。 相似文献
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T. Beda 《Journal of Polymer Science.Polymer Physics》2007,45(13):1713-1732
A constitutive phenomenological model completing the Gent‐Thomas concept is carried out to formulate laws governing the hyperelastic behavior of incompressible rubber materials. It is shown that the phenomenological Gent‐Thomas model (1958) and the constrained chain model (1992) give similar precise results at small to moderate deformation. On the other hand, comparisons of the outcome of the proposed model with that of the molecular model from the combined concepts of Flory‐Erman and Boyce‐Arruda (2000), and with those of the phenomenological models of Ogden (1982), Yeoh‐Fleming (1997), Pucci‐Saccomandi (2002) and Beda (2005) are made. Residual inconveniences raised by attractive continuum models in rubber elasticity literature have been successfully overcome. Results from both the statistical and phenomenological mechanics concepts are compared with the data of some useful classical materials (rubbers of Treloar, Rivlin‐Saunders, Pak‐Flory and Yeoh‐Fleming). The results permit one to see salient equivalence of the two theories for a more reliable prediction of stress‐stretch response for all states of any mode of deformation. A complete and exhaustive analysis of the Mooney plot that combines small and very large extension‐compression has been quite essential in assessing the validity of models. A method of identification of material parameters is presented and data of the simple tension suffice for the determination of the parameter values. It is shown that the ordinary identification procedures, such as the usual least squares, a very much used numerical method in materials investigation, can be unsuitable in some cases of hyperelastic modeling. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 1713–1732, 2007 相似文献
4.
以陆地棉(冀棉20)下胚轴为外植体,在含KT0.3mg/L和2,4-D0.08mg/L的MS培养基上,诱导产生了胚性愈伤组织,继代培养不需要添加激素,胚性愈伤组织即分化为胚状体,并萌发出小芽,胚状体萌发过程与合子胚相似,但出现了一些畸形胚,解剖学方法观察发现,这些畸形胚的维管束系统不明显或形态异常,以农杆菌介导法对冀棉20胚状体及胚性愈伤组织进行了雪花莲凝集素(GNA)基因的遗传转化条件探索,并以GUS基域瞬间进行检测加以证实,得出农杆菌的浓度为O.D600=0.8时,浸染时间为5min,共培养时间为3d,筛选培养基中卡那霉素浓度为75mg/L,对冀棉20胚状体及胚性愈伤组织转化是较为适合的。 相似文献
5.
Ken Tachibana Ken Takeda Masahiko Shiraishi 《Proceedings of the Japan Academy. Series B, Physical and biological sciences》2004,80(7):342
It is well-recognized that DNA methylation and histone modifications play critical roles in epigenetic regulation of gene activity through the alteration of chromatin structure. Recent studies have shown that in a subset of cancer cells, the silencing of the human E-cadherin (CDH1) gene is associated with hypermethylation of the CpG island. However, the associated molecular mechanism remains unclear. To understand the mechanism, we have investigated the alteration of CpG island methylation and histone modifications during the reactivation of the CDH1 gene by treatment with 5-aza-2′-deoxycytidine (5-aza-dC). Although the CDH1 gene expression was recovered by treatment with 5-aza-dC in a liver cancer cell line Li21, the methylation status of the entire CpG island and acetylation and methylation status of associated histones were not significantly altered. These results demonstrate that the silenced CDH1 gene can be reactivated without apparent alteration of histone modification or CpG island methylation. 相似文献
6.
The interaction between gene activation and cellular activity has recently emerged as a critical aspect of brain behavior, but the dynamics of networks incorporating these interactions are poorly understood. An interesting phenomena arises when the genetic activation oscillates endogenously and a network of such cells synchronize to a coherent rhythm, such as is the case with the suprachiasmatic nucleus. To explain this synchronization, we propose a model in which a mRNA/protein expression cycle drives neurons electrical activity, and synaptic activation shifts the phase of the protein rhythm. Using lattice networks, we demonstrate that these interactions are sufficient to generate coherent oscillation. © 2006 Wiley Periodicals, Inc. Complexity 12: 67–72, 2006 相似文献
7.
根据Taura综合征病毒(TSV)基因组,设计特异性引物,从感染病毒组织中提取组织总RNA后扩增,分别将3个主要结构蛋白基因VP1、VP2和VP3克隆到pGEM TEasyVector.与表达载体连接后,导入大肠杆菌中诱导表达,并纯化目的蛋白.诱导表达的融合蛋白分子量分别为54.2×103、43×103和57.1×103,在变性条件下过柱纯化VP1和VP2,一次可以纯化10mg以上纯度较高的蛋白. 相似文献
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Zhi-Hui Su Munehiro Okamoto Osamu Tominaga Katsumi Akita Noboru Kashiwai Y?ki Imura Tooru Ojika Yoshiyuki Nagahata Syozo Osawa 《Proceedings of the Japan Academy. Series B, Physical and biological sciences》2006,82(7):232-250
An approach to deduce the mechanism of stabilization of the hybrid-derived populations in the Ohomopterus ground beetles has been made by comparative studies on the phylogenetic trees of the mitochondrial and nuclear DNA. A phylogenetic tree based on the internal transcribed spacer (ITS) of nuclear ribosomal gene roughly reflects the relations of morphological species group, while mitochondrial NADH dehydrogenase subunit 5 (ND5) gene shows a considerable different topology on the tree; there exist several geographically-linked lineages, most of which consist of more than one species. These results suggest that the replacement of mitochondria has occurred widely in the Ohomopterus species. In most cases, hybridization is unidirectional, i.e., the species A (♂) hybridized with another species B (♀) and not vice versa, with accompanied replacement of mitochondria of A by those of B. The results also suggest that partial or complete occupation of the distribution territory by a hybrid-derived morphological species. The morphological appearance of the resultant hybrid-derivatives are recognized as that of the original species A. Emergence of a morphological new species from a hybrid-derived population has been exemplified. 相似文献
10.
在缺失了3'LTR U3区内病毒的启动子/增强子序列的逆转录病毒载体pLXSNd中,用血管内皮生长因子受体KDR的特异性启动子调控了TNFa在血管内细胞ECV304中的靶向表达。将构建的载体pLXSN-TNFa,pLXSNd-KDRp-TNFa和空载体pLXSN用PA317细胞包装后获得重组病毒,并用重组病毒分别感染NIH3T3细胞和ECV304细胞,培养物上清的ELISA结果证明,KDR启动子指导的TNFa在KDR阳性细胞ECV304中的表达量为在KDR阴性细胞NIH3T3中的表达量的8倍;而TR指导的TNFa在这两种细胞中的表达无明显差异,实现了TNFa在血管内皮细胞中的靶向表达,这可能为肿瘤基因治疗提供新途径。 相似文献