全文获取类型
收费全文 | 2218篇 |
免费 | 224篇 |
国内免费 | 1篇 |
出版年
2022年 | 18篇 |
2021年 | 32篇 |
2020年 | 14篇 |
2019年 | 15篇 |
2018年 | 25篇 |
2017年 | 13篇 |
2016年 | 45篇 |
2015年 | 72篇 |
2014年 | 70篇 |
2013年 | 87篇 |
2012年 | 113篇 |
2011年 | 123篇 |
2010年 | 79篇 |
2009年 | 75篇 |
2008年 | 111篇 |
2007年 | 114篇 |
2006年 | 104篇 |
2005年 | 90篇 |
2004年 | 90篇 |
2003年 | 80篇 |
2002年 | 57篇 |
2001年 | 65篇 |
2000年 | 52篇 |
1999年 | 59篇 |
1998年 | 31篇 |
1997年 | 13篇 |
1996年 | 34篇 |
1995年 | 31篇 |
1994年 | 29篇 |
1993年 | 32篇 |
1992年 | 36篇 |
1991年 | 32篇 |
1990年 | 37篇 |
1989年 | 24篇 |
1988年 | 30篇 |
1987年 | 31篇 |
1986年 | 19篇 |
1985年 | 23篇 |
1984年 | 11篇 |
1983年 | 13篇 |
1981年 | 11篇 |
1980年 | 13篇 |
1979年 | 12篇 |
1978年 | 9篇 |
1975年 | 11篇 |
1974年 | 9篇 |
1973年 | 10篇 |
1970年 | 13篇 |
1969年 | 9篇 |
1927年 | 9篇 |
排序方式: 共有2443条查询结果,搜索用时 234 毫秒
31.
Localization of phycoerythrin at the lumenal surface of the thylakoid membrane in Rhodomonas lens 总被引:2,自引:0,他引:2
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The thylakoids of cryptomonads are unique in that their lumens are filled with an electron-dense substance postulated to be phycobiliprotein. In this study, we used an antiserum against phycoerythrin (PE) 545 of Rhodomonas lens (gift of R. MacColl, New York State Department of Health, Albany, NY) and protein A-gold immunoelectron microscopy to localize this light-harvesting protein in cryptomonad cells. In sections of whole cells of R. lens labeled with anti-PE 545, the gold particles were not uniformly distributed over the dense thylakoid lumens as expected, but instead were preferentially localized either over or adjacent to the thylakoid membranes. A similar pattern of labeling was observed in cell sections labeled with two different antisera against PE 566 from Cryptomonas ovata. To determine whether PE is localized on the outer or inner side of the membrane, chloroplast fragments were isolated from cells fixed in dilute glutaraldehyde and labeled in vitro with anti-PE 545 followed by protein A-small gold. These thylakoid preparations were then fixed in glutaraldehyde followed by osmium tetroxide, embedded in Spurr, and sections were labeled with anti-PE 545 followed by protein A-large gold. Small gold particles were found only at the broken edges of the thylakoids, associated with the dense material on the lumenal surface of the membrane, whereas large gold particles were distributed along the entire length of the thylakoid membrane. We conclude that PE is located inside the thylakoids of R. lens in close association with the lumenal surface of the thylakoid membrane. 相似文献
32.
λ-Glutamylcysteine synthetase in higher plants: catalytic properties and subcellular localization 总被引:1,自引:0,他引:1
λ-Glutamylcysteine synthetase activity (EC 6.3.2.2) was analysed in Sephacryl S-200 eluents of extracts from cell suspension
cultures ofNicotiana tabacum L. cv. Samsun by determination of λ-glutamylcysteine as its monobromobimane derivative. The enzyme has a relative molecular
mass (Mr) of 60000 and exhibits maximal activity at pH 8 (50% at pH 7.0 and pH9.0) and an absolute requirement for Mg2+. With 0.2mM Cd2+ or Zn2+, enzyme activity was reduced by 35% and 19%, respectively. Treatment with 5 mM dithioerythritol led to a heavy loss of activity
and to dissociation into subunits (Mr 34000). Buthionine sulfoximine andl-methionine-sulfoximine, known as potent inhibitors of λ-glutamylcysteine synthetase from mammalian cells, were found to be
effective inhibitors of the plant enzyme too. The apparent Km values forl-glutamate,l-cysteine, and α-aminobutyrate were, respectively, 10.4mM, 0.19 mM, and 6.36 mM. The enzyme was completely inhibited by glutathione
(Ki=0.42 mM). The data indicate that the rate of glutathione synthesis in vivo may be influenced substantially by the concentration
of cysteine and glutamate and may be further regulated by feedback inhibition of λ-glutamylcysteine synthetase by glutathione
itself. λ-Glutamylcysteine synthetase is, like glutathione synthetase, localized in chloroplasts as well as in the cytoplasm.
Chloroplasts fromPisum sativum L. isolated on a Percoll gradient contained about 72% of the λ-glutamylcysteine synthetase activity in leaf cells and 48%
of the total glutathione synthetase activity. In chloroplasts ofSpinacia oleracea L. about 61% of the total λ-glutamylcysteine synthetase activity of the cells were found and 58% of the total glutathione
synthetase activity. These results indicate that glutathione synthesis can take place in at least two compartments of the
plant cell.
Dedicated to Professor A. Prison on the occasion of his 80th birthday 相似文献
33.
Haemophilia B (sixth edition): a database of point mutations and short additions and deletions. 总被引:2,自引:1,他引:1
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
F Giannelli P M Green S S Sommer M C Poon M Ludwig R Schwaab P H Reitsma M Goossens A Yoshioka G G Brownlee 《Nucleic acids research》1996,24(1):103-118
The sixth edition of the haemophilia B database lists in easily accessible form all known factor IX mutations due to small changes (base substitutions and short additions and/or deletions of <30 bp) identified in haemophilia B patients. The 1380 patient entries are ordered by the nucleotide number of their mutation. Where known, details are given on factor IX activity, factor IX antigen in circulation and origin of mutation. References to published mutations are given and the laboratories generating the data are indicated. 相似文献
34.
35.
In culture, Azorhizobium caulinodans used at least four terminal oxidases, cytochrome aa3 (cytaa3), cytd, cyto, and a second a-type cytochrome, which together mediated general, respiratory electron (e-) transport to O2. To genetically dissect physiological roles for these various terminal oxidases, corresponding Azorhizobium apocytochrome genes were cloned, and three cytaa3 mutants, a cytd mutant, and a cytaa3, cytd double mutant were constructed by reverse genetics. These cytochrome oxidase mutants were tested for growth, oxidase activities, and N2 fixation properties both in culture and in symbiosis with the host plant Sesbania rostrata. The cytaa3 mutants grew normally, fixed N2 normally, and remained fully able to oxidize general respiratory e- donors (NADH, succinate) which utilize a cytc-dependent oxidase. By difference spectroscopy, a second, a-type cytochrome was detected in the cytaa3 mutants. This alternative a-type cytochrome (Amax = 610 nm) was also present in the wild type but was masked by bona fide cytaa3 (Amax = 605 nm). In late exponential-phase cultures, the cytaa3 mutants induced a new, membrane-bound, CO-binding cytc550, which also might serve as a cytc oxidase (a fifth terminal oxidase). The cloned Azorhizobium cytaa3 genes were strongly expressed during exponential growth but were deactivated prior to onset of stationary phase. Azorhizobium cytd mutants showed 40% lower N2 fixation rates in culture and in planta, but aerobic growth rates were wild type. The cytaa3, cytd double mutant showed 70% lower N2 fixation rates in planta. Pleiotropic cytc mutants were isolated by screening for strains unable to use N,N,N',N'-tetramethyl-p-phenylenediamine as a respiratory e- donor. These mutants synthesized no detectable cytc, excreted coproporphyrin, grew normally in aerobic minimal medium, grew poorly in rich medium, and fixed N2 poorly both in culture and in planta. Therefore, while aerobic growth was sustained by quinol oxidases alone, N2 fixation required cytc oxidase activities. Assuming that the terminal oxidases function as do their homologs in other bacteria, Azorhizobium respiration simultaneously employs both quinol and cytc oxidases. Because Azorhizobium terminal oxidase mutants were able to reformulate their terminal oxidase mix and grow more or less normally in aerobic culture, these terminal oxidases are somewhat degenerate. Its extensive terminal oxidase repertoire might allow Azorhizobium spp. to flourish in wide-ranging O2 environments. 相似文献
36.
Soil columns in which the root system was divided into threeequal layers, each 24 cm in diameter and 33 cm high were usedto examine the influence of drying different proportions ofthe root system on the water relations, gas exchange and abscisicacid (ABA) concentration of lupin (Lupinus cosentinii Guss.cv. Eregulla) leaves. The treatments imposed were (i) all threelayers adequately watered (control), (ii) the upper layer unwateredwith the remaining layers kept adequately watered, (iii) thetwo upper layers unwatered with the basal layer kept adequatelywatered, (iv) all three layers unwatered. The treatments wereapplied at 56 d after sowing (DAS), and continued for 21 d inthe treatment in which the three layers were dried and for 36d in the other three treatments. After 21 d, the soil matricpotential in the layers that were unwatered had decreased toemdash 1.3MPa, compared to - 0.03 MPa in the adequately-wateredlayers. Within 8 d of cessation of watering, plants with the entireroot system in drying soil had significantly lower stomatalconductances, lower rates of net photosynthesis, and higherleaf ABA contents than did adequately-watered plants. Whilethe leaf osmotic potential decreased within 8 d of cessationof watering, the leaf water potential did not change for thefirst 15 d after water was withheld. After withholding waterfrom all layers, the shoot dry matter was 63% lower than thatin the adequately-watered plants. In the two partially-droughtedtreatments, 17% and 48% of the root length was subjected todrying. Compared to the adequately-watered plants, drying upto 50% of the root system for 36 d, in the two partially-droughtedtreatments, did not reduce stomatal conductance, net photosynthesis,or plant growth. Similarly, there was no significant effecton leaf water potential or osmotic potential. When either theupper or upper and middle layers of soil were dried, the ABAcontent of the leaves for most of the drying period was slightly,but not significantly, higher than in leaves of the adequately-wateredplants. The results suggest that lupins with a well-established rootsystem can utilize localized supplies of available soil waterto maintain leaf gas exchange despite appreciable portions ofthe root system being in dry soil. In contrast to other studies,the results also suggest that when only a portion of the soilvolume is dry and adequate water is available in the wet zone,root signals do not influence stomatal conductance and leafgas exchange of lupin. Key words: Abscisic acid, gas exchange, lupins, split-roots, water deficit 相似文献
37.
R. Landmann U. Keilholz C. Scheibenbogen M. Brockhaus H. Gallati H. Denz M. Bargetzi C. Ludwig 《Cancer immunology, immunotherapy : CII》1994,38(2):113-118
Eleven metastatic cancer patients were studied during three different regimens of immunotherapy with interleukin-2 (IL-2) and/or interferon (IFN): group A received 4 days of IL-2 i.a. infusion (n=3), group B IFN s.c. during 5 days (n=4), followed on day 3 by 5 days of a continuous IL-2 i.v. infusion, and group C had 4 days of IL-2 i.v. infusion together with s.c. IFN on days 1 and 4 (n=4). Soluble tumor necrosis factor receptors (sTNFR) p55 and p75 and TNF concentrations in serum were analyzed before therapy and daily during 8 days of the first therapy cycle. sTNFR was measured by radioimmunoassay. sTNFR p55 increased in all patient groups from a baseline value of 5.2±0.9 ng/ml to a maximum of 13.6±1.2 ng/ml by days 3–4 (P=0.003). sTNFR p75 increased from 7.6±1.1 ng/ml to peak values of 30.1±2.6 ng/ml in groups A and B (P=0.02). In group C the sTNFR p75 response was weak (NS). In group B, the increase of both p55 and p75 occurred only after addition of IL-2 to IFN. TNF increased weakly during treatment with IFN alone (group B); it rose strongly during IL-2 and the combined treatment (groups A-C) from 8±2 pg/ml to 115±13 pg/ml (P=0.003). In group B, it reached the maximum 24 h after addition of IL-2 to IFN and decreased thereafter. there was a significant relationship between TNF and sTNFR p55 or sTNFR p75 in groups A and C, (P=0.001), but not in group B. Group C was also investigated during the third therapy cycle. The increase of sTNFR p75 was stronger (P=0.01) and that of TNF weaker than in the first cycle; the sTNFR p55 response was similar in both cycles. In conclusion sTNFR p55 and p75 are rapidly induced during IL-2 and IL-2+IFN treatment, the increase of sTNF receptors parallels or exceeds that of TNF and may influence the immunomodulatory effects of TNF during cytokine therapy. 相似文献
38.
16S rDNA sequence and phylogenetic position of an uncultivated spirochete from the hindgut of the termite Mastotermes darwiniensis Froggatt 总被引:3,自引:0,他引:3
Abstract We have analyzed the 16S rDNA sequence and the phylogenetic position of an uncultivated spirochete from the hindgut contents of the Australian termite Mastotermes darwiniensis Froggatt. The 16S rRNA genes of bacteria from the hindgut contents of Mastotermes darwiniensis were amplified by polymerase chain reaction. The amplification products were cloned and sequenced. The sequences were compared to known homologous primary structures. Two of the clones (MDS1 and MDS3) had an insert of 1498 nucleotides showing typical signatures of spirochete 16S rRNA sequences. The sequences of the two clones were most similar to the 16S rRNA sequence of Spirochaeta stenostrepta (89.8%) and Treponema sp. strain H1 (90.7%). Phylogenetical analysis positioned the hindgut spirochete sequence with that of the free-living anaerobic Spirochaeta stenostrepta and Treponema sp. strain H1 as its nearest relatives within the cluster of the spirochetes. We conclude that the analyzed SSU rDNA sequences originate from a spirochete related to the genus Treponema . It is possibly one of the uncultivated unique spirochetes symbiotic in termite hindguts. 相似文献
39.
Adherent recombinant BHK cells were cultivated at temperatures between 30 and 37°C. Batch and repeated-batch-cultivations in a 2-litre bioreactor showed a significant influence on metabolism and cell growth. The low-temperature-cultivations showed a lower growth rate and a lower glucose consumption rate and, therefore, less lactate production. On the other hand, the maximum cell density and productivity seemed not to be affected by the temperature reduction. 相似文献
40.
WIP1, a wound-inducible gene from maize with homology to Bowman-Birk proteinase inhibitors 总被引:6,自引:0,他引:6
We have cloned and sequenced a wound-inducible cDNA clone designated WIP1 (for wound-induced protein) from maize coleoptiles. It was isolated by differential screening of a cDNA library prepared from excised maize coleoptile segments. The deduced amino acid sequence predicts a secretory, cysteine-rich protein of 102 residues with a calculated molecular mass of 11 kDa and a typical N-terminal signal sequence. The protein has about 30% identity with various Bowman-Birk type proteinase inhibitors. Most interestingly, it is novel in that it is double-headed with exclusive specificity for chymotrypsin. WIP1 is strongly wound-induced in contrast to other members of the Bowman-Birk proteinase inhibitor family, which occur in seeds and are regulated during development. The response is fast, similar to defenceinduced genes, and measurable as early as 30 min after wounding. Induction can also be evoked in the intact coleoptiles and the signal is systemically transmitted in the coleoptile to adjacent regions of the wounded area. Isolation and analysis of the corresponding genomic clone reveals that WIP1 contains an intron of 90 nucleotides. 相似文献