全文获取类型
收费全文 | 2285篇 |
免费 | 268篇 |
出版年
2021年 | 16篇 |
2019年 | 18篇 |
2018年 | 24篇 |
2016年 | 37篇 |
2015年 | 70篇 |
2014年 | 64篇 |
2013年 | 88篇 |
2012年 | 104篇 |
2011年 | 99篇 |
2010年 | 50篇 |
2009年 | 53篇 |
2008年 | 99篇 |
2007年 | 99篇 |
2006年 | 89篇 |
2005年 | 86篇 |
2004年 | 98篇 |
2003年 | 69篇 |
2002年 | 76篇 |
2001年 | 88篇 |
2000年 | 65篇 |
1999年 | 58篇 |
1998年 | 36篇 |
1997年 | 36篇 |
1996年 | 26篇 |
1995年 | 26篇 |
1994年 | 22篇 |
1993年 | 22篇 |
1992年 | 39篇 |
1991年 | 33篇 |
1990年 | 54篇 |
1989年 | 49篇 |
1988年 | 42篇 |
1987年 | 32篇 |
1986年 | 41篇 |
1985年 | 36篇 |
1984年 | 29篇 |
1983年 | 29篇 |
1982年 | 19篇 |
1981年 | 20篇 |
1979年 | 29篇 |
1978年 | 30篇 |
1977年 | 21篇 |
1976年 | 32篇 |
1975年 | 23篇 |
1974年 | 23篇 |
1973年 | 26篇 |
1972年 | 24篇 |
1971年 | 17篇 |
1970年 | 21篇 |
1969年 | 15篇 |
排序方式: 共有2553条查询结果,搜索用时 15 毫秒
61.
A splicing-dependent regulatory mechanism that detects translation signals. 总被引:32,自引:3,他引:29 下载免费PDF全文
Premature termination codons (PTCs) can cause the decay of mRNAs in the nuclear fraction of mammalian cells. This enigmatic nuclear response is of interest because it suggests that translation signals do not restrict their effect to the cytoplasm, where fully assembled ribosomes reside. Here we examined the molecular mechanism for this putative nuclear response by using the T-cell receptor-beta (TCR-beta) gene, which acquires PTCs as a result of programmed rearrangements that occur during normal thymic ontogeny. We found that PTCs had little or no measurable effect on TCR-beta pre-mRNA levels, but they sharply depressed TCR-beta mature mRNA levels in the nuclear fraction of stably transfected cells. A PTC split by an intron was able to trigger the down-regulatory response, implying that PTC recognition occurs after an mRNA is at least partially spliced. However, intron deletion and addition studies demonstrated that a PTC must be followed by at least one functional (spliceable) intron to depress mRNA levels. One explanation for this downstream intron-dependence is that cytoplasmic ribosomes adjacent to nuclear pores scan mRNAs still undergoing splicing as they emerge from the nucleus. We found this explanation to be unlikely because PTCs only 8 or 10 nt upstream of a terminal intron down-regulated mRNA levels, even though this distance is too short to permit PTC recognition in the cytoplasm prior to the splicing of the downstream intron in the nucleus. Collectively, the results suggest that nonsense codon recognition may occur in the nucleus. 相似文献
62.
PCR analysis of oilseed rape cultivars (Brassica napus L. ssp. oleifera) using 5′ -anchored simple sequence repeat (SSR) primers 总被引:8,自引:0,他引:8
Y. M. Charters A. Robertson M. J. Wilkinson G. Ramsay 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,92(3-4):442-447
Primers complementary to simple sequence repeats (SSRs) and with variable three-base anchors at their 5 end, were used in PCR analyses to compare pooled DNA samples from various Brassica napus and B. rapa cultivars. Amplification products were resolved on polyacrylamide gels and detected by silver-nitrate staining. The resulting banding patterns were highly repeatable between replicate PCRs. Two of the primers produced polymorphisms at 33 and 23 band positions, respectively, and could each discriminate 16 of the 20 cultivars studied. Combined use of both primers allowed all 20 cultivars to be distinguished. The UPGMA dendrogram, based on the cultivar banding profiles, demonstrated clustering on the basis of winter/spring growth habit, high/low glucosinolate content, and cultivar origin (i.e. the breeder involved). Intracultivar polymorphism was investigated using a minimum of ten individuals for each cultivar and was found to vary considerably between cultivars. It is concluded that anchored SSR-PCR analysis is a highly informative and reproducible method for fingerprinting oilseed rape populations, but that intra-cultivar variation should be investigated before using banding profiles from pooled samples for the identification of individuals. 相似文献
63.
Cold stress proteins induced in Listeria monocytogenes in response to temperature downshock and growth at low temperatures. 总被引:3,自引:1,他引:2 下载免费PDF全文
Listeria monocytogenes is a food-borne pathogen with the ability to grow at refrigerator temperatures. Twelve cold shock proteins (Csps) with apparent M(r)s of 48,600, 41,000, 21,800, 21,100, 19,700, 19,200, 18,800, 18,800, 17,200, 15,500, 14,500, and 14,400 were induced by cold shocking L. monocytogenes 10403S from 37 to 5 degrees C, as revealed by labeling with L-[35S]methionine followed by two-dimensional gel electrophoresis. Strain SLCC53 showed a similar response. Cold acclimation proteins were observed in cultures of strain 10403S growing at 5 degrees C, and four of these proteins, with apparent M(r)s 48,000, 21,100, 19,700, and 18,800, were also Csps. Two cold-sensitive transposon-induced mutants were labeled less efficiently than the parent strain, but the Csp response of the mutant examined was very similar to that of the parent strain. 相似文献
64.
65.
Gern N. M. Huijberts Hetty van der Wal Clare Wilkinson Gerrit Eggink 《Biotechnology Techniques》1994,8(3):187-192
Summary The accuracy and reproducibility of the gas-chromatographic method for the analysis of PHB and PHA in whole cells of Alcaligenes eutrophus H16 and Pseudomonas putida KT2442 were determined. It was found that for analysis of PHA the methanolysis time in the assay had to be increased to 4 h. Accuracy of the PHB and PHA assay were 0.018 mg and 0.304 mg respectively, based on estimation of the measurement error. 相似文献
66.
Graham J. Moore Renee C. Ganter John M. Matsoukas John Hondrelis George Agelis Klemoenis Barlos Scott Wilkinson John Sandall Patrick Fowler 《Journal of molecular recognition : JMR》1994,7(4):251-256
A triad of interacting group (TyrOH? His$ \underline\ominus$O2C) in angiotensin II (ANG II) has been postulated to create the tyrosinate anion pharmacophore (tyanophore) responsible for receptor activation/triggering (Biochim. Biophys. Acta 1991, 1065, 21). In the present study we investigated the effects on bioactivity of substituting the Tyr4 residue in [Sar1]ANG II with other anionic or electronegative amino acids, and with a number of aromatic amino acids lacking a hydroxyl group. [Sar1 Nva(δ-OH)4]ANG II, [Sar1 Nva(δ-OCH3)4]ANG II, [Sar1 Met4]ANG II, [Sar1 Gln4]ANG II, [Sar1 Glu4]ANG II and [Sar1 DL -Alg4]ANG II had agonist activities in the rat isolated uterus assay of 4, 3, 19, 10, > 0.1 and > 0.1%, respectively, of that of ANG II. [Sar1 Nal4]ANG II, [Sar1 Pal4]ANG II, [Sar1 DL -Phg(4′-F)4]ANG II, [Sar1 Phe(4′-F)4]ANG II, [Sar1 Phe(F5)4]ANG II and [Sar1 His4]ANG II had agonist activities of 4.5, 7, < 0.1, 0.2, 1 and 0.6%, respectively. All peptides investigated were devoid of measurable antagonist activity except [Sar1] Phe(4′-F)4 ANG II (pA2 = 7.7). These findings illustrate that anionic or electronegative aliphatic side chains replacing tyrosinate at position 4 can partially activate the angiotension receptor. For ANG II analogues containing an aromatic amino acid other than Tyr at position 4, ligand binding and agonist activity are not dependent on the electronegativity or dipole moment of the aromatic ring, or on the ability of the 4′ ring substituent to accept a proton. Modelling based on ab initio calculations of aromatic ring multipoles illustrate that the apparent binding affinity (PA2) of ANG II analogues is associated with a perpendicular electrostatic interaction of the position 4 aromatic ring with a receptor-based group. In addition, intramolecular interactions providing for the conformation of the ligand as it approaches its receptor appear to have a role in determining agonist vs antagonist activity. 相似文献
67.
Mutant plants defective in the assimilation of nitrate can be selected by their resistance to the herbicide chlorate. In Arabidopsis thaliana, mutations at any one of nine distinct loci confer chlorate resistance. Only one of the CHL genes, CHL3, has been shown genetically to be a nitrate reductase (NR) structural gene (NIA2) even though two NR genes (NIA1 and NIA2) have been cloned from the Arabidopsis genome. Plants in which the NIA2 gene has been deleted retain only 10% of the wildtype shoot NR activity and grow normally with nitrate as the sole nitrogen source. Using mutagenized seeds from the NIA2 deletion mutant and a modified chlorate selection protocol, we have identified the first mutation in the NIA1 NR structural gene. nia1, nia2 double mutants have only 0.5% of wild-type shoot NR activity and display very poor growth on media with nitrate as the only form of nitrogen. The nial-1 mutation is a single nucleotide substitution that converts an alanine to a threonine in a highly conserved region of the molybdenum cofactor-binding domain of the NR protein. These results show that the NIA1 gene encodes a functional NR protein that contributes to the assimilation of nitrate in Arabidopsis. 相似文献
68.
The amino-terminal one-third of pseudorabies virus glycoprotein gIII contains a functional attachment domain, but this domain is not required for the efficient penetration of Vero cells. 总被引:7,自引:6,他引:1 下载免费PDF全文
We have examined the attachment and penetration phenotypes of several glycoprotein gIII mutants of pseudorabies virus (PRV) and have identified the first one-third of gIII as a region that mediates efficient virus attachment to PK15 and Vero cells. This portion of gIII, amino acids 25 through 157 of the wild-type sequence, appeared to support attachment by binding to heparinlike molecules on cell surfaces. Virions containing the first one-third of gIII were sensitive to heparin competition and showed greatly reduced infectivity on cells treated with heparinase. PRV virions lacking the first one-third of the mature glycoprotein exhibited only residual binding to cells if challenged by vigorous washing with phosphate-buffered saline at 2 h postinfection at 4 degrees C. This residual binding was resistant to heparin competition, and strains lacking the first one-third of gIII were able to infect cells treated with heparinase as effectively as untreated cells. When we determined the penetration phenotypes for each strain, we found that gIII-mediated virus attachment was necessary for timely penetration of PK15 cells but remarkably was not required for efficient virus penetration of Vero cells. Moreover, wild-type PRV was actually prohibited from rapid penetration of Vero cells by a gIII-heparan sulfate interaction. Our results indicate that initial virus binding to heparan sulfate via glycoprotein gIII is not required for efficient PRV infection of all cell types and may in fact be detrimental in some instances. 相似文献
69.
T cell receptor-beta mRNA splicing: regulation of unusual splicing intermediates. 总被引:10,自引:3,他引:7 下载免费PDF全文
L Qian L Theodor M Carter M N Vu A W Sasaki M F Wilkinson 《Molecular and cellular biology》1993,13(3):1686-1696
70.
Structural and serological characterisation of an O-specific polysaccharide from Serratia plymuthica 总被引:4,自引:0,他引:4
Abstract The surface polysaccharides of a strain of Serratia plymuthica were characterised and shown to consist of a linear, acidic galactoglucomannan as well as a major and a minor neutral galactan. Immunoblotting results demonstrated cross-reactions between this strain and others with similar galactans ( S. marcescens O16 and O20, Klebsiella O1, and Pasteurella haemolytica T4 and T10). 相似文献