全文获取类型
收费全文 | 75638篇 |
免费 | 7249篇 |
国内免费 | 4665篇 |
专业分类
87552篇 |
出版年
2024年 | 145篇 |
2023年 | 827篇 |
2022年 | 1954篇 |
2021年 | 3231篇 |
2020年 | 2239篇 |
2019年 | 2686篇 |
2018年 | 2634篇 |
2017年 | 2056篇 |
2016年 | 2911篇 |
2015年 | 4077篇 |
2014年 | 4928篇 |
2013年 | 5084篇 |
2012年 | 5984篇 |
2011年 | 5588篇 |
2010年 | 3358篇 |
2009年 | 3076篇 |
2008年 | 3538篇 |
2007年 | 3219篇 |
2006年 | 2843篇 |
2005年 | 2393篇 |
2004年 | 1977篇 |
2003年 | 1888篇 |
2002年 | 1523篇 |
2001年 | 3025篇 |
2000年 | 2880篇 |
1999年 | 2219篇 |
1998年 | 901篇 |
1997年 | 884篇 |
1996年 | 846篇 |
1995年 | 762篇 |
1994年 | 661篇 |
1993年 | 533篇 |
1992年 | 1125篇 |
1991年 | 891篇 |
1990年 | 765篇 |
1989年 | 628篇 |
1988年 | 506篇 |
1987年 | 421篇 |
1986年 | 344篇 |
1985年 | 280篇 |
1984年 | 200篇 |
1983年 | 187篇 |
1982年 | 125篇 |
1981年 | 86篇 |
1980年 | 78篇 |
1979年 | 93篇 |
1976年 | 76篇 |
1974年 | 78篇 |
1973年 | 75篇 |
1972年 | 76篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
991.
992.
Molecular diffusion into ferritin: pathways, temperature dependence, incubation time, and concentration effects 总被引:2,自引:0,他引:2 下载免费PDF全文
The detailed kinetics of permeation and effusion of small nitroxide spin probe radicals with the protein shells of horse spleen ferritin (HoSF) and human H-chain ferritin (HuHF) and a 3-fold channel variant D131H+E134H of HuHF were studied by electron paramagnetic resonance spectroscopy and gel permeation chromatography under a variety of experimental conditions. The results confirm that the permeation of molecular species of 7-9-A diameter into ferritin is a charge selective process and that the threefold channels are the likely pathways for entry into the protein. Studies with holoHoSF show that increased temperature increases the rates of penetration and effusion and also increases the concentration of positively charged spin probe accumulated within the protein in excess of that in the external solution. The interior of HoSF is much more accessible to small molecules at physiological temperature of approximately 40 degrees C than at room temperature. The large activation energy of 63-67 kJ/mol measured for the effusion/penetration and the small diffusion coefficient, D approximately 5 x 10(-22) m(2)/s at 20 degrees C, corresponding to a time of approximately 60 min for traversing the protein shell, is consistent with the kinetics of diffusion being largely controlled by the restrictive porosity of the protein itself. An inverse dependence of the first-order rate constant for effusion out of the protein channel on the incubation time used for radical penetration into the protein is attributed to increased binding of the radical within the funnel-shaped channel. 相似文献
993.
Jian-Rao Hu Mei Liu Da-Hui Wang Yan-Jun Hu Fu-Qing Tan Wan-Xi Yang 《Molecular biology reports》2013,40(12):6645-6655
The member of the kinesin-14 subfamily, KIFC1, is a carboxyl-terminal motor protein that plays an important role in the elongation of nucleus and acrosome biogenesis during the spermiogenesis of mammals. Here, we had cloned and sequenced the cDNA of a mammalian KIFC1 homologue (termed ec-KIFC1) from the total RNA of the testis of the reptile Eumeces chinensis. The full-length sequence was 2,339 bp that contained a 216 bp 5′-untranslated region (5′UTR), a 194 bp 3′-untranslated region (3′UTR) and a 1,929 bp open reading frame that encoded a special protein of 643 amino acids (aa). The calculated molecular weight of the putative ec-KIFC1 was 71 kDa and its estimated isoelectric point was 9.47. The putative ec-KIFC1 protein owns a tail domain from 1 to 116 aa, a stalk domain from 117 to 291 aa and a conserved carboxyl motor domain from 292 to 642 aa. Protein alignment demonstrated that ec-KIFC1 had 45.6, 42.8, 44.6, 36.9, 43.7, 46.4, 45.1, 55.6 and 49.8 % identity with its homologues in Mus musculus, Salmo salar, Danio rerio, Eriocheir sinensis, Rattus norvegicus, Homo sapiens, Bos taurus, Gallus gallus and Xenopus laevis, respectively. Tissue expression analysis showed the presence of ovary, heart, liver, intestine, oviduct, testis and muscle. The phylogenetic tree revealed that ec-KIFC1 was more closely related to vertebrate KIFC1 than to invertebrate KIFC1. In situ hybridization showed that the ec-KIFC1 mRNA was localized in the periphery of the nuclear membrane and the center of the nucleus in early spermatids. In mid spermatids, the ec-KIFC1 had abundant expression in the center of nucleus, and was expressed in the tail and the anterior part of spermatids. In the late spermatid, the nucleus gradually became elongated, and the ec-KIFC1 mRNA signal was still centralized in the nucleus. In mature spermatids, the signal of the ec-KIFC1 gradually became weak, and was mainly located at the tail of spermatids. Therefore, the ec-KIFC1 probably plays a critical role in the spermatogenesis of E. chinensis. 相似文献
994.
A simple and sensitive assay for 1-aminocyclopropane-1-carboxylic acid 总被引:99,自引:0,他引:99
A simple, rapid, and sensitive method for the quantitative determination of 1-amino-cyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene in plant tissues, is described. The assay is based on the liberation of ethylene from ACC with NaOCl in the presence of Hg2+; ethylene is assayed by gas chromatography. The yield is normally 80% and can be determined by internal standards. The method is quite specific and can detect as little as 5 pmol of ACC. 相似文献
995.
Rhizobitoxine is a phytotoxin synthesized by some strains of the legume symbiont genus Bradyrhizobium and the plant pathogen Pseudomonas andropogonis. We demonstrate here a new enzymatic assay which is 100-fold more sensitive than previous assays and can detect as little as 1.0 pmol of rhizobitoxine. The assay is based on the inhibition of Salmonella typhimurium beta-cystathionase by rhizobitoxine. Interestingly, beta-cystathionase from Bradyrhizobium japonicum is insensitive to rhizobitoxine at concentrations lower than 75 microM. 相似文献
996.
The relationship between human cytomegalovirus infection and atherosclerosis development 总被引:14,自引:0,他引:14
Transport characteristics of essential trace elements as zinc, copper, selenium and iron have been studied in maternal–fetal direction in normal pregnancies, using in vitro perfusion of human placental lobules. Solutions of trace elements corresponding to twice the physiological concentrations were injected (100 l bolus) into the maternal arterial perfusate. Serial perfusate samples were collected every 30 sec from venous outflows for a study period of 5 min. Concentrations of these trace elements and their transport kinetics were determined. Transport fractions (TF) of zinc, copper, selenium and iron averaged 0.21, 0.49, 0.55 and 0.10% of maternal load respectively. Other parameters such as area under the curve, clearance, elimination constant, absorption and elimination rates showed some significant differences between the various elements. Copper and selenium appear to be transported passively in maternal–fetal direction, while for iron and zinc, role of active transport for transfer across the human placental membrane cannot be discounted. We speculate that alterations in copper: zinc TR50 (transport rate for 50% efflux) and TF ratios could serve as useful indicators for assessing placental transport status of these essential elements in complicated pregnancy states. 相似文献
997.
Development and evaluation of an enzyme-linked immunosorbent assay with recombinant SAG2 for diagnosis of Toxoplasma gondii infection in cats 总被引:7,自引:0,他引:7
Huang X Xuan X Kimbita EN Battur B Miyazawa T Fukumoto S Mishima M Makala LH Suzuki H Sugimoto C Nagasawa H Fujisaki K Mikami T Igarashi I 《The Journal of parasitology》2002,88(4):804-807
Cats are pivotal in the transmission of Toxoplasma gondii. To develop a sensitive and specific serodiagnostic method for feline toxoplasmosis, surface antigen 2 (SAG2) of T. gondii was expressed in Escherichia coli and its diagnostic potential evaluated in an enzyme-linked immunosorbent assay (ELISA). The ELISA with recombinant SAG2 (rSAG2) was able to differentiate very clearly between sera from cats experimentally infected with T. gondii and sera from normal cats. Serum samples collected from domestic cats in Japan were investigated by the ELISA, and the results were compared with those of a commercially available latex agglutination test (LAT) kit. Of the 192 samples screened, 42 (21.9%) were positive by ELISA. Among the 42 ELISA-positive samples, 39 were positive by LAT. There was a significant correlation between ELISA and LAT titers. All the 150 ELISA-negative samples were negative by LAT. These results indicate that the ELISA with rSAG2 expressed in E. coli should be a useful method for detection of T. gondii infection in cats. 相似文献
998.
[目的]生物质的利用是当前生物技术研究的一个热点.本小组分离到一株高效降解纤维素球毛壳菌(Chaetomium globosum)NK102,本文拟探索研究此菌的纤维素酶表达系统并寻找影响酶基因表达的关键因素.[方法]通过对NK102测序,本文界定了球毛壳菌NK102的主要纤维素酶编码基因,使用数字基因表达谱升级版(RNA-Seq)的方法得到纤维素酶基因的表达差异,然后观察了营养、物理条件下纤维素酶基因表达和酶活性变化的情况.[结果]发现随着培养时间的延长,纤维素酶基因整体上表达量升高.在所选基因中,外切葡聚糖酶、纤维二糖脱氢酶和内切葡聚糖酶基因(cbh1,cdh和egl1)的表达量最高.糖代谢的负调控因子ACE I和CreA的随时间表达量均降低,而Hap2/3/5复合体的表达量反而升高.之后检测了不同碳源培养基对纤维素酶基因表达量和酶活性的影响,发现葡萄糖为强阻遏因子,纤维二糖为其诱导物,而山梨醇没有影响.特别是,我们发现光照也影响纤维素酶基因的表达,黑暗条件明显抑制酶基因的表达.[结论]转录组学的方法可以初步探索纤维素酶表达的规律,酶基因的表达受到营养、物理条件的影响.本研究为揭示球毛壳菌降解纤维素分子机理和阐释生物质糖代谢途径提供了有用参考. 相似文献
999.
Removal of the RecA C-terminus results in a conformational change in the RecA-DNA filament 总被引:1,自引:0,他引:1
The Escherichia coli RecA protein catalyzes homologous recombination of DNA molecules, and the active form of the protein is a helical polymer that it forms around DNA. Previous image analysis of electron micrographs has revealed the RecA protein to be organized into two domains or lobes within the RecA-DNA filament. We have now been able to show that a small modification of the RecA protein by proteolysis results in a significant shift in the internal mass in the RecA filament. We have cleaved approximately 18 residues from the C-terminus of the RecA protein, producing a roughly 36K MW RecA core protein that binds DNA and polymerizes normally. A three-dimensional reconstruction of this complex has been computed, and has been compared with a previous reconstruction of the intact protein. The main difference is consistent with a 15 A outward movement of the lobe that was at an inner radius in the wild-type protein. These observations yield additional evidence about the conformational flexibility of the RecA filament, and will aid in understanding the structural mechanics and dynamics of the RecA filament. 相似文献
1000.
衡山青冈栎群落结构及物种多样性研究 总被引:1,自引:0,他引:1
利用样方法进行野外群落调查,共设计了15个样方,每个样方大小为20m×5 m,对衡山青冈栎(Cyclobalanopsis glauca)群落的种类组成、外貌、结构特征及物种多样性进行了研究.结果表明:在1 500 m2样地中,共有维管束植物26科37属46种,其中优势科为壳斗科、山茶科和蔷薇科等;该群落主要是以青冈栎为主的单优种群落,生活型谱以高位芽植物占绝对优势,叶级谱以中、小型叶为优势的特点,群落季相不明显;群落分层较明显,乔木层树种很少,灌木层种类较丰富,对群落物种多样性具有较大影响;物种多样性分析显示该群落多样性指数偏低,其多样性复杂程度由高到低的顺序为灌木层、藤本层、乔木层、草本层. 相似文献