全文获取类型
收费全文 | 27458篇 |
免费 | 1538篇 |
国内免费 | 4172篇 |
出版年
2024年 | 50篇 |
2023年 | 380篇 |
2022年 | 599篇 |
2021年 | 769篇 |
2020年 | 719篇 |
2019年 | 913篇 |
2018年 | 745篇 |
2017年 | 801篇 |
2016年 | 909篇 |
2015年 | 1049篇 |
2014年 | 1313篇 |
2013年 | 1946篇 |
2012年 | 1127篇 |
2011年 | 1300篇 |
2010年 | 1062篇 |
2009年 | 1453篇 |
2008年 | 1525篇 |
2007年 | 1632篇 |
2006年 | 1536篇 |
2005年 | 1484篇 |
2004年 | 1359篇 |
2003年 | 1305篇 |
2002年 | 1082篇 |
2001年 | 858篇 |
2000年 | 712篇 |
1999年 | 672篇 |
1998年 | 690篇 |
1997年 | 564篇 |
1996年 | 527篇 |
1995年 | 538篇 |
1994年 | 509篇 |
1993年 | 406篇 |
1992年 | 384篇 |
1991年 | 291篇 |
1990年 | 243篇 |
1989年 | 230篇 |
1988年 | 212篇 |
1987年 | 165篇 |
1986年 | 122篇 |
1985年 | 163篇 |
1984年 | 191篇 |
1983年 | 122篇 |
1982年 | 125篇 |
1981年 | 79篇 |
1980年 | 60篇 |
1979年 | 62篇 |
1978年 | 47篇 |
1977年 | 26篇 |
1976年 | 40篇 |
1974年 | 26篇 |
排序方式: 共有10000条查询结果,搜索用时 812 毫秒
91.
Tomato (Lycopersicon esculentum Mill. cv. Moneymaker) plants have been wounded to induce the accumulation of proteinase-inhibitor proteins (PI proteins) at the local site of injury and systemically in unwounded tissues. To determine the range of genes affected in the wound-response, polysomal mRNA has been isolated from the damaged leaves and from systemically responding leaves over a time-course of 2, 4, 10 and 24 h after wounding. Changes in the pattern of 35S-translation products indicate that the events that occur at the local wound-site are different from those that occur systemically, both with respect to the number of genes that are regulated and the timing of their regulation. In order to compare the effects of wounding and an endogenous systemic signal generated at the wound-site with those of elicitor (proteinase-inhibitor-inducing factor, PIIF) treatment of excised plants, polysomal mRNA has also been isolated from leaves of plants over a time-course of 2, 4, 10 and 24 h after PIIF-treatment. Changes in the pattern of 35S-translation products indicates that the events induced by PIIF resemble those induced by mechanical injury, rather than those induced by the endogenous systemic signal.Abbreviations IFF
isoelectric focussing
- PI proteins
proteinase inhibitor proteins
- PIIF
proteinase-inhibitor-inducing factor
- ssRubisco
small subunit of ribulose-1,5-bisphosphate carboxylase 相似文献
92.
Komandoor E. Achyuthan Ann Mary Robin Bhaermani Charles S. Greenberg 《Molecular and cellular biochemistry》1989,85(1):57-65
Calcium ions are crucial for expression of transglutaminase activity. Although lanthanides have been reported to substitute for calcium in a variety of protein functions, they did not replace the calcium requirement during transglutaminase activity measurements. Furthermore, lanthanides strongly inhibited purified liver transglutaminase activity using either casein or fibrinogen as substrates. Terbium (III) inhibition of transglutaminase-catalyzed putrescine incorporation into casein was not reversed by the presence of 10–200 fold molar excess of calcium ions (Ki for Tb(III)=60 µM). Conformational changes in purified liver transglutaminase upon Tb(III) binding were evident from a biphasic effect of Tb(III) on transglutaminase binding to fibrin. Low concentrations of Tb(III) (1 µM to 10 µM inhibited the binding of transglutaminase to fibrin, whereas higher concentrations (20 µM to 100 µM promoted binding. Conformational changes in purified liver transglutaminase consequent to Tb(III) binding were also demonstrated by fluorescence spectroscopy due to Forster energy transfer. Fluorescence emission was stable to the presence of 200 mM NaCl and 100 mM CaCl2 only partially quenched emission. Purified liver transglutaminase strongly bound to Tb(III)-Chelating Sepharose beads and binding could not be disrupted by 100 mM CaCl2 solution. Our data suggest that Tb(III)-induced conformational changes in transglutaminase are responsible for the observed effects on enzyme structure and function. The potential applications of Tb(III)-transglutaminase interactions in elucidating the structure-function relationships of liver transglutaminase are discussed. 相似文献
93.
94.
The chalcone synthase multigene family of Petunia hybrida (V30): differential,light-regulated expression during flower development and UV light induction 总被引:11,自引:0,他引:11
We have analysed the expression of the 8–10 members of the gene family encoding the flavonoid biosynthetic enzyme chalcone synthase (CHS) from Petunia hybrida. During normal plant development only two members of the gene family (CHS-A and CHS-J) are expressed. Their expression is restricted to floral tissues mainly. About 90% of the total CHS mRNA pool is transcribed from CHS-A, wheares CHS-J delivers about 10% in flower corolla, tube and anthers. Expression of CHS-A and CHS-J during flower development is coordinated and (red) light-dependent. In young seedlings and cell suspension cultures expression of CHS-A and CHS-J can be induced with UV light. In addition to CHS-A and CHS-J, expression of another two CHS genes (CHS-B and CHS-G) is induced in young seedlings by UV light, albeit at a low level. In contrast to CHS genes from Leguminoseae, Petunia CHS genes are not inducible by phytopathogen-derived elicitors. Expression of CHS-A and CHS-J is reduced to a similar extent in a regulatory CHS mutant, Petunia hybrida Red Star, suggesting that both genes are regulated by the same trans-acting factors. Comparison of the promoter sequences of CHS-A and CHS-J reveals some striking homologies, which might represent cis-acting regulatory sequences. 相似文献
95.
Kees W. Rodenburg Marcel J. A. de Groot Rob A. Schilperoort Paul J. J. Hooykaas 《Plant molecular biology》1989,13(6):711-719
In relation to the question which DNA form (single- or double-stranded) is transferred by Agrobacterium tumefaciens to plant cells, we studied the behaviour of single-stranded DNA, as compared to double-stranded DNA, when it is introduced into plant protoplasts by electroporation. To this end, we cloned a construct with a plant NPTII gene as well as a CAT gene in the M13 vectors tg130 and tg131. We found that both complementary single-stranded molecules gave rise to substantial CAT activity in plant protoplasts, suggesting that single-stranded DNA is converted into double-stranded DNA by the plant cell replication machinery. Unexpectedly, we found that single-stranded DNA leads to a 3–10 fold higher frequency of stable transformation (selection for kanamycin resistance) than double-stranded DNA. These results indicate that the use of single-stranded DNA might be considered in experiments in which optimal transformation frequencies are needed, e.g. with protoplasts form recalcitrant plant species.Abbreviations ss
single-stranded
- ds
double-stranded
- CAT
chloramphenicol acetyl transferase
- NPTII
neomycin phosphotransferase II
- RT
room temperature 相似文献
96.
97.
D. K. Dougall 《Plant Cell, Tissue and Organ Culture》1989,18(1):105-112
Approximately half of the subclones examined from one clone of the wild carrot cell culture WC63-1-9-1 accumulated dihydroquercetin in the culture medium. The amount of dihydroquercetin accumulated in the medium varied with the subclone used, the size of the inoculum, the medium used and the time of sampling.Dedicated to Dr. Friedrich Constabel on the occasion of his 60th birthday 相似文献
98.
Green cybrids with a new nucleus-chloroplast combination cannot be selected after protoplast fusion in the intersubfamilial Nicotiana-Solanum combination. As an approach to overcome the supposed plastomegenome incompatibility, a partial plastome transfer by genetic recombination has been considered. After fusions of protoplasts of a light-sensitive Nicotiana tabacum (tobacco) plastome mutant and lethally irradiated protoplasts of wild-type Solanum tuberosum (potato), a single green colony was recovered among 2.5×104 colonies. The regenerated plants had tobacco-like (although abnormal) morphology, but were normally green, and sensitive to tentoxin, demonstrating chloroplast markers of the potato parent. Restriction enzyme analysis of the chloroplast DNA (cpDNA) revealed recombinant, nonparental patterns. A comparison with physical maps of the parental cpDNA demonstrated the presence of a considerable part of the potato plastome flanked by tobacco-specific regions. This potacco plastome proved to be stable in backcross and backfusion experiments, and normally functional in the presence solely of N. tabacum nucleus. 相似文献
99.
1. The working hypothesis that neuropeptide gene expression in a neuron is an indicator of that neuron's physiological activity is discussed. 2. Representative examples from the literature are presented to support the hypothesis. 3. Further, we discuss the regulation of expression of two opioid peptides, preproenkephalin and preprodynorphin, in laminae I and II of the spinal cord and in nucleus caudalis of the trigeminal nuclear complex, where they may play a role in pain modulation. 4. The expression of the opioid peptide genes can be induced by both painful and nonnoxious stimuli in neurons in time-dependent and sensory-specific fashions. 相似文献
100.
A variety of bacterial O-polysaccharides were covalently linked to enzymes and it was demonstrated with three discrete monoclonal antibodies that enzyme-glycoconjugates function as convenient labelled antigens in direct enzyme immunoassays, particularly competitive assays that quantify bacterial O-antigens. Two strategies, each involving reductive amination, were used to couple O-polysaccharides to enzymes, while retaining high enzymic activity. Reduction of the Schiff base formed between, 1,3-diaminopropane and the terminal reducing ketodeoxyoctanoic acid (KDO) residue present in the majority of the lipopolysaccharide (LPS) core domains, following mild acid removal of Lipid A, offered the most direct route to mono-aminated polysaccharide. Alternatively, mild periodate oxidation of KDO and heptose residues generated multiple aldehyde targets for Schiff base formation, without affecting the O-antigenic determinant. Hetero- and homobifunctional coupling reagents, sulphosuccinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate and discuccinimidyl suberate, activated polysaccharide for coupling to enzymes at amino and sulphydryl sites and produced conjugates that retained at least 95% of the original enzymic activity. The most suitable enzyme conjugates, especially for competitive inhibition EIA were those bearing one polysaccharide chain, and these were easily prepared from horse-radish peroxidase. Although the extent of conjugation of activated polysaccharide to -galactosidase and alkaline phosphatase could be controlled by reaction stoichiometry, the use of these enzymes was a less effective utilization of valuable antigen and enzyme.Issued as NRCC No. 31634 相似文献