全文获取类型
收费全文 | 7737篇 |
免费 | 676篇 |
国内免费 | 874篇 |
出版年
2024年 | 40篇 |
2023年 | 174篇 |
2022年 | 341篇 |
2021年 | 454篇 |
2020年 | 335篇 |
2019年 | 466篇 |
2018年 | 387篇 |
2017年 | 271篇 |
2016年 | 387篇 |
2015年 | 521篇 |
2014年 | 592篇 |
2013年 | 623篇 |
2012年 | 683篇 |
2011年 | 585篇 |
2010年 | 359篇 |
2009年 | 332篇 |
2008年 | 364篇 |
2007年 | 315篇 |
2006年 | 295篇 |
2005年 | 208篇 |
2004年 | 238篇 |
2003年 | 206篇 |
2002年 | 173篇 |
2001年 | 157篇 |
2000年 | 121篇 |
1999年 | 93篇 |
1998年 | 74篇 |
1997年 | 54篇 |
1996年 | 80篇 |
1995年 | 64篇 |
1994年 | 50篇 |
1993年 | 27篇 |
1992年 | 42篇 |
1991年 | 32篇 |
1990年 | 31篇 |
1989年 | 21篇 |
1988年 | 24篇 |
1987年 | 16篇 |
1986年 | 12篇 |
1985年 | 18篇 |
1984年 | 6篇 |
1983年 | 7篇 |
1982年 | 5篇 |
1981年 | 4篇 |
排序方式: 共有9287条查询结果,搜索用时 15 毫秒
71.
The NSR1 gene encodes a protein that specifically binds nuclear localization sequences and has two RNA recognition motifs 总被引:29,自引:6,他引:23 下载免费PDF全文
We previously identified a protein (p67) in the yeast, Saccharomyces cerevisiae, that specifically recognizes nuclear localization sequences. We report here the partial purification of p67, and the isolation, sequencing, and disruption of the gene (NSR1) encoding this protein. p67 was purified using an affinity column conjugated with a peptide containing the histone H2B nuclear localization sequence from yeast. Using antibodies against p67 we have cloned the gene for this protein. The protein encoded by the NSR1 gene recognizes the wild-type H2B nuclear localization sequence, but does not recognize a mutant H2B sequence that is incompetent for nuclear localization in vivo. Interestingly, the NSR1 protein has two RNA recognition motifs, as well as an acidic NH2 terminus containing a series of serine clusters, and a basic COOH terminus containing arg-gly repeats. We have confirmed the nuclear localization of p67 by immunofluorescence and found that a restricted portion of the nucleus is highlighted. We have also shown that NSR1 (p67) is required for normal cell growth. 相似文献
72.
转移及非转移肿瘤移植后615小鼠血液流变学变化的研究 总被引:1,自引:0,他引:1
血道高转移瘤株FC、淋巴合并血道高转移瘤株U14、淋巴道高转移瘤株H22、非转移瘤株P615分别接种于336只纯系近交615小鼠.不同时间取血并处死动物,进行组织学及血液流变学检查.将转移瘤发展过程分为潜伏期、侵袭期、转移早、中、晚期,非转移瘤发展过程分为潜优期、增殖期、囊腔形成期及中心坏死期.本实验结果显示,不同转移能力及途径肿瘤发展的不同时期血液流变学变化规律不同,因而表明肿瘤侵袭、转移与血液流变学变化之间存在互为因果的紧密关系.其临床诊断及治疗意义被讨论. 相似文献
73.
以Lineweave-Burk plot双倒数作图法测得该酶对底物S-腺苷酰甲硫氨酸(SAM)的K_m=7.69×10~(-6)mol/L,在1mmol/LS-腺苷酰高半胱氨酸(SAH)存在下,Ki=7.33×10~(-4)mol/L,两条直线相交于纵轴,证明SAH是该酶的竞争性抑制剂。该酶最适pH为7.8,对热不稳定。同时还测定了该酶对不同DNA底物的专一性及盐浓度、代谢相关物’两价阳离子、某些酸根等对该酶调节性质的影响。以碘代乙酰胺修饰该酶的SH基’及用二硫苏糖醇(DTT)和巯基乙醇(MSH)保护该酶SH基所作的实验表明SH基是该酶活性中心所必需的,用高效液相色谱(HPLC)法证明该酶所甲基化的碱基为刘氏小球菌(M·L、DNA)分子中的胞嘧啶,且求得甲基化30min后所得甲基化水平为2.39%。同时也证明当用该酶将λDNA甲基化后,可使BamHI限制性核酸内切酶对甲基化后的λDNA丧失切割作用。 相似文献
74.
Triterpenoid saponins from Clinopodium polycephalum. 总被引:4,自引:0,他引:4
A new triterpenoid saponin, clinopodiside A, has been isolated from Clinopodium polycephalum. Its structure was established by spectroscopic methods and X-ray diffraction analysis as 3-O-beta-D-glucopyranosyl(1----6)-[ beta-D-glucopyranosyl (1----4)]-beta-D-glucopyranosyl-olean-11,13(18)-diene-3 beta,16 beta, 23,28-tetrol. 相似文献
75.
Total in vitro maturation of the Saccharomyces cerevisiae a-factor lipopeptide mating pheromone 总被引:4,自引:0,他引:4
S Marcus G A Caldwell C B Xue F Naider J M Becker 《Biochemical and biophysical research communications》1990,172(3):1310-1316
The a-factor mating pheromone, produced by Saccharomyces cerevisiae a haploid cells, is post-translationally modified in a manner analogous to that of the ras proto-oncogene product. A consensus C-terminal amino acid sequence, -CAAX (C is cysteine, A is aliphatic amino acid, and X is any amino acid), is the target of these modifications, which include isoprenylation (essential for Ras function), proteolysis of the -AAX sequence, and carboxy methyl esterification. Recently, the RAM/DPR1 gene product was shown to be a component of the activity responsible for isoprenylation of both Ras and a-factor. In this report, we present an in vitro assay which not only detects a-factor isoprenylation, but also proteolysis and carboxy methyl esterification, and directly demonstrates, biochemically, the order of these processing events. This a-factor maturation assay may prove useful for screening agents which block any of the steps involved in the post-translational modification of the a-factor and Ras -CAAX sequences. Such agents would be potential anti-Ras-related cancer therapeutic drugs. 相似文献
76.
db—cAMP对转化细胞钙调素基因表达与细胞骨架的影响 总被引:5,自引:0,他引:5
We have demonstrated that the distribution of microtubules (MT), microfilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c-fos enhanced in the transformed C3 H10 T1/2 cells. After treatment with 1 mM db-cAMP for 1 hr. and 2 hrs., there was an early and rapidly reduced in gene expression of calmodulin and c-fos respectively. After db-cAMP treatment for 4-5 days, the number of Capping cells of ConA binding decreased significantly and the cell surface microvilli decreased also. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed in G1 phase, we have found that the db-cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, microfilaments and fibronectin were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the inhibition of proliferation, alteration of phenotype and recovery of cytoskeleton in transformed cells after treatment with db-cAMP are related to the inhibition of gene expression of calmodulin. 相似文献
77.
78.
To identify sources of high potency antigens for use in serodiagnosis, aqueous-soluble egg antigens from Schistosoma japonicum were extracted with Dulbecco's phosphate-buffered saline. Residual particulates were solubilized with Tris-buffered 8 M urea, yielding a urea-soluble egg antigen fraction. The urea-soluble fraction was further fractionated with Bio Gel A50m and QAE-Sephadex. All fractions were quantitatively assayed for their specific antigenic activities against serum specimens from infected rabbits by the single-tube enzyme-linked immunosorbent assay (k-ELISA). In antigen rate-limiting conditions, the urea-soluble particulate fractions were more antigenically active than the aqueous-soluble fraction. In antigen-excess and antibody-limiting assay conditions, the ideal conditions for serologic assays, the urea-derived antigens also showed superior activities against sera from infected humans. Analysis by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on gradient gels revealed numerous low molecular weight protein bands in the aqueous-soluble fraction, whereas the urea-soluble fractions appeared to be much simpler with the majority of their proteins concentrated in one or two high molecular weight bands (greater than or equal to 200 kdaltons). Electro-transfer blots of the SDS-PAGE onto nitrocellulose papers and subsequent visualization of antigens by enzyme-linked immunoabsorbence confirmed these findings. The above data suggest that the urea-soluble fraction of S. japonicum eggs is antigenically active and has potential use in the development of a diagnostic reagent. 相似文献
79.
黄土区不同恢复年限草地群落生物量及根冠比对氮添加的响应 总被引:1,自引:0,他引:1
大气氮沉降增加作为全球变化的主要环境问题之一,已引发人们的广泛关注,持续的氮沉降对草地生态系统的组成、结构和功能产生重要影响。为深入了解草地恢复进程中群落生物量和根冠比对氮沉降的响应,以黄土区3个不同恢复年限(初期12a、中期28a和后期37a)的天然草地为研究对象,通过设置6个氮添加水平,CK (0)、N1(2.34g m-2a-1)、N2(4.67g m-2a-1)、N3(9.34g m-2a-1)、N4(18.68g m-2a-1)、N5(37.35g m-2a-1)来测定草地群落地上生物量、地下生物量和总生物量,并计算根冠比和氮响应效率(NRE)。结果表明:(1)地上生物量在恢复中期最大,随氮添加梯度增加,地上生物量在恢复初期和恢复后期呈不显著上升趋势,对氮添加表现为非线性的正响应(ΔNRE>0),在恢复中期呈不显著下降趋势,对氮添加表现为非线性的负响应(ΔNRE<0)。(2)群落地下生物量对氮添加无显著响应,总生物量只有在恢复后期的N4添加水平下,与对照存在显著差异。(3)根冠比在恢复初期时,N3添加水平下显著高于对照和其他氮添加水平,其余恢复年限对氮添加无显著响应。综上所述,通过分析比较黄土区不同恢复年限草地群落的地上、地下及总生物量和根冠比对氮添加的响应。建议对该区域开展试点实验,实行适应性草地管理,如进行两年一次刈割或轻度放牧(2只羊/hm2),来探寻更科学有效的管理措施,使草地实现系统性恢复,进而满足生态系统容量和社会需求的变化。 相似文献