全文获取类型
收费全文 | 14491篇 |
免费 | 792篇 |
国内免费 | 502篇 |
专业分类
15785篇 |
出版年
2024年 | 18篇 |
2023年 | 158篇 |
2022年 | 259篇 |
2021年 | 307篇 |
2020年 | 310篇 |
2019年 | 344篇 |
2018年 | 386篇 |
2017年 | 275篇 |
2016年 | 297篇 |
2015年 | 411篇 |
2014年 | 603篇 |
2013年 | 826篇 |
2012年 | 399篇 |
2011年 | 504篇 |
2010年 | 451篇 |
2009年 | 559篇 |
2008年 | 579篇 |
2007年 | 615篇 |
2006年 | 559篇 |
2005年 | 532篇 |
2004年 | 492篇 |
2003年 | 471篇 |
2002年 | 417篇 |
2001年 | 325篇 |
2000年 | 297篇 |
1999年 | 281篇 |
1998年 | 278篇 |
1997年 | 257篇 |
1996年 | 223篇 |
1995年 | 289篇 |
1994年 | 266篇 |
1993年 | 260篇 |
1992年 | 246篇 |
1991年 | 233篇 |
1990年 | 211篇 |
1989年 | 234篇 |
1988年 | 238篇 |
1987年 | 208篇 |
1986年 | 168篇 |
1985年 | 233篇 |
1984年 | 353篇 |
1983年 | 246篇 |
1982年 | 301篇 |
1981年 | 239篇 |
1980年 | 206篇 |
1979年 | 192篇 |
1978年 | 66篇 |
1977年 | 51篇 |
1976年 | 44篇 |
1973年 | 17篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
51.
Stanislaw Ulaszewski Elisabetta Balzi André Goffeau 《Molecular & general genetics : MGG》1987,207(1):38-46
Summary In the yeast Saccharomyces cerevisiae, the pma1 mutations confers vanadate-resistance to H+-ATPase activity when measured in isolated plasma membranes. In vivo, the growth of pma1 mutants is resistant to Dio-9, ethidium bromide and guanidine derivatives. This phenotype was used to man the pma1 mutation adjacent to LEU1 gene on chromosome VII. From a cosmid library of a wild-type Saccharomyces cerevisiae genome, a large 30 kb DNA fragment was isolated by complementation of a leu1-pma1 double mutant. A 5 kb HindIII fragment was subcloned and it restored both Leu+ and Pma+ phenotypes after integrative transformation. The restriction map of the 5 kb HindIII fragment and Southern blot analysis reveal that the cloned fragment contains the entire structural gene for the plasma membrane ATPase and the 5 end of the adjacent LEU1 gene. The pma1 mutation conferring vanadate-resistance is thus located in the structural gene for the plasma membrane ATPase.Publication no 2456 from the Biology Directorate of the Commission of European Communities 相似文献
52.
Monoclonal antibodies generated by immunization with a plasma-membrane preparation from suspension-cultured cells of Nicotiana glutinosa L. were used in combination with fluoresceinor rhodamine-labeled goat anti-mouse immunoglobulins to identify heterokaryons in protoplast fusion procedures. Antibody labeling did not inhibit callus formation nor plantlet regeneration. The antibodies are non-invasive and surface labeling provides clear optical discrimination of true heterokaryons from unfused aggregates as well as from parental protoplasts and homokaryons. Labeling is stable throughout fusion and hence by pre-labeling parental protoplast populations the strategy is both versatile and of general applicability. 相似文献
53.
Summary Fluorescein or rhodamine conjugates of seventeen different lectins were tested for their ability to label the plasma membrane of live plant protoplasts. During the investigation, a strong effect of calcium was observed on the binding of several lectins to protoplasts derived from suspension cultured rose cells (Rosa sp. Paul's Scarlet). The binding of these lectins was increased by elevating the calcium concentration from 1 to 10 mM in the buffer. Other divalent cations had variable, but similar, effects on lectin binding. The mechanism of this effect appeared to involve the protoplast surface rather than the lectins. Although the cell wall-degrading enzymes used to isolate protoplasts had generally no effect on lectin binding, one clear exception was observed. Binding ofArachis hypogaea agglutinin was markedly reduced on protoplasts isolated with Driselase as compared to protoplasts isolated with a combination of Cellulysin and Pectolyase Y-23. Although most of the lectins that labeled protoplasts derived from cultured rose cells or from corn root cortex (Zea mays L. WF9 × Mo17) had specificities for galactose or N-acetylgalactosamine, some differences in protoplast labeling between lectins of the same saccharide specificity were observed. Two different analyses of the interaction betweenRicinus communis agglutinin and rose protoplasts showed that binding was cooperative with an apparent association constant of 7.2 × 105M–1 or 9.8 × 105M–1 with a maximum of approximately 108 lectin molecules bound per protoplast. Treatment of protoplasts with glycosidases which hydrolyze either N- or O-glycosidic linkages of glycoproteins slightly enhanced labeling of protoplasts byRicinus communis agglutinin. Interpretation of these results are discussed.Abbreviations MPR
medium, minimal organic medium (Nothnagel andLyon 1986)
- APA
Abrus precatorius agglutinin
- CSA
Cytisus sessilifolius agglutinin
- ECA
Erythrina cristagalli agglutinin
- GS-I
Griffonia simplicifolia agglutinin
- LcH
Lens culinarus agglutinin
- PNA
Arachis hypogaea agglutinin
- SBA
Glycine max agglutinin
- VAA
Viscum album agglutinin
- VFA
Vicia faba agglutinin
- WGA
Triticum vulgaris agglutinin
- Con A
Canavalia ensiformis agglutinin
- HPA
Helix pomatia agglutinin
- TPA
Tetragonolobus purpureas agglutinin
- RCA
Ricinus communis agglutinin
- DBA
Dolichos biflorus agglutinin
- SJA
Sophora japonica agglutinin
- BPA
Bauhinia purpurea agglutinin
- FITC
fluorescein isothiocyanate
- Ga1NAc
N-acetylgalactosamine
- FDA
fluorescein diacetate
- 2-O-Me-D-Fuc
2-O-methyl-D-fucose
Parts of the work presented here are also submitted in partial fulfillment of requirements for the Ph.D. degree. 相似文献
54.
S. K. Podder A. Chakraborti K. Vijayalakshmi P. Lalit Kumar Singh 《Journal of biosciences》1987,11(1-4):495-502
To examine how surface Potential controls the reactivity of glycoconjugates at cell surface, the interaction of galactose-sPecific
lectinse.g. peanut agglutinin,Ricinus cummunis agglutinin with liPosomes bearing asialo GM1 were studied in the Presence of varying amount of ganglioside mixture, GMn. The Presence of 5% GMn causes comPlete slowing down of PreciPitin reaction and thereby make carbohydrate moiety of asialo GM1 comPletely inaccessiblei.e. ‘cryPtic’. In contrast the Presence of 1–2% GMn enhances the aPParent rate and amPlitude of the PreciPitin reaction as surface Potential becomes more negative. The relevance
of the findings has been discussed in relation to the exPression and involvement of the cell-surface sialic acid residues
during develoPment and differentiation. 相似文献
55.
Lawrence G. Palmer 《The Journal of membrane biology》1987,96(2):97-106
Epithelial Na channels are apparently pore-forming membrane proteins which conduct Na much better than any other biologically abundant ion. The conductance to Na can be 100 to 1000 times higher than that to K. The only other ions that can readily get through this channel are protons and Li. Small organic cations cannot pass through the channel, and water may also be impermeant. The selectivity properties of epithelial Na channels appear to be determined by at least three factors: A high field-strength anionic site, most likely a carboxyl residue of glutamic or aspartic acid residues on the channel protein, probably accounts for the high conductance through these channels of Na and Li and to the low conductance of K, Rb and Cs. A restriction in the size of the pore at its narrowest point probably accounts for the low conductance of organic cations as well as the possible exclusion of water molecules. The outer mouth of the channel appears to be negatively charged and may control access to the region of highest selectivity and may serve as a preliminary selectivity filter, attracting cations over anions. These conclusions are illustrated by the cartoon of the channel in Fig. 3. This picture is obviously both fanciful and simplified, but its general points will hopefully be testable. It leaves open a number of important questions, including: does amiloride block the channel by binding within the outer mouth? what does the inner mouth of the channel look like, and does this part of the channel contribute to selectivity? and what, if any, are the interactions between the features of the channel that impart selectivity and those that control the regulation of the channel by hormonal and other factors? 相似文献
56.
Summary The presence of a Ca2+ channel in the plasmalemma of tonoplast-freeNitellopsis obtusa cells was demonstrated and its characteristics were studied using current- and voltage-clamp techniques. A long-lasting inward membrane current (I
m
), recorded using a step voltage clamp, consisted of a single component without time-dependent inactivation. Increasing either [Ca2+]
o
or [Cl–]
o
1) enhanced the maximum amplitude of inwardI
m
((I
m
)
p
) and 2) shifted the peak voltage ((V
m
)
p
) at(I
m
)
p
to more positive values under ramp-shaped voltage clamping and 3) depolarized the peak value of action potentials. This behavior is consistent with predictions based on the Nernst equation for Ca2+ but not for Cl–. DIDS (4,4-diisothiocyano-2,2-disulfonic acid stilbene) did not suppress(I
m
)
p
in tonoplast-free cells, in contrast with its effect on normal cells. La3+ and nifedipine blocked(I
m
)
p
irreversibly. On the other hand, Ca2+ channel agonist, BAY K 8644 irreversibly enhanced(I
m
)
p
. Both Sr2+ influx and K+ efflux increased upon excitation. The charge carried by Sr2+ influx was compensated for by K+ efflux. It is concluded that only the Ca2+ channel is activated during plasmalemma excitation in tonoplast-free cells. In terms of the magnitude of(I
m
)
p
, Sr2+ could replace Ca2+, but Mn2+, Mg2+ and Ba2+ could not. External pH affected(I
m
)
p
and the membrane conductance (g
m
) at(I
m
)
p
((g
m
)
p
). Increasing the external ionic strength caused increases in both(I
m
)
p
and(g
m
)
p
, and shifted(V
m
)
p
to positive values. At the same time, Sr2+ influx increased. Thus Ca2+ channel activation seems to be enhanced by increasing external ionic strength. The possible involvement of surface potential is discussed. 相似文献
57.
Summary The mitochondrial outer membrane contains voltagegated channels called VDAC that are responsible for the flux of metabolic substrates and metal ions across this membrane. The addition of micromolar quantities of aluminum chloride to phospholipid membranes containing VDAC channels greatly inhibits the voltage dependence of the channels' permeability. The channels remain in their high conducting (open) state even at high membrane potentials. An analysis of the change in the voltage-dependence parameters revealed that the steepness of the voltage dependence decreased while the voltage needed to close half the channels increased. The energy difference between the open and closed states in the absence of an applied potential did not change. Therefore, the results are consistent with aluminum neutralizing the voltage sensor of the channel. pH shift experiments showed that positively charged aluminum species in solution were not involved. The active form was identified as being either (or both) the aluminum hydroxide or the tetrahydroxoaluminate form. Both of these could reasonably be expected to neutralize a positively charged voltage sensor. Aluminum had no detectable effect of either single-channel conductance or selectivity, indicating that the sensor is probably not located in the channel proper and is distinct from the selectivity filter. 相似文献
58.
Summary Phloridzin-insensitive, Na+-independentd-glucose uptake into isolated small intestinal epithelial cells was shown to be only partially inhibited by trypsin treatment (maximum 20%). In contrast, chymotrypsin almost completely abolished hexose transport. Basolateral membrane vesicles prepared from rat small intestine by a Percoll® gradient procedure showed almost identical susceptibility to treatment by these proteolytic enzymes, indicating that the vesicles are predominantly oriented outside-out. These vesicles with a known orientation were employed to investigate the kinetics of transport in both directions across the membrane. Uptake data (i.e. movement into the cell) showed aK
t of 48mm and aV
max of 1.14 nmol glucose/mg membrane protein/sec. Efflux data (exit from the cell) showed a lowerK
t of 23mm and aV
max of 0.20 nmol glucose/mg protein/sec.d-glucose uptake into these vesicles was found to be sodium independent and could be inhibited by cytochalasin B. TheK
t for cytochalasin B as an inhibitor of glucose transport was 0.11 m and theK
D for binding to the carrier was 0.08 m.d-glucose-sensitive binding of cytochalasin B to the membrane preparation was maximized withl- andd-glucose concentrations of 1.25m. Scatchard plots of the binding data indicated that these membranes have a binding site density of 8.3 pmol/mg membrane protein. These results indicate that the Na+-independent glucose transporter in the intestinal basolateral membrane is functionally and chemically asymmetric. There is an outward-facing chymotrypsin-sensitive site, and theK
t for efflux from the cell is smaller than that for entry. These characteristics would tend to favor movement of glucose from the cell towards the bloodstream. 相似文献
59.
Summary Brush border membrane vesicles were prepared from mussel gills using differential and sucrose density gradient centrifugation. These vesicles contained both the maximal Na+-dependent alanine transport activity found in the gradient and the maximal activities of -glutamyl transpeptidase and alkaline phosphatase. Electron micrographs showed closed vesicles of approximately 0.1–0.5 m diameter. Transport experiments using these vesicles demonstrated a transient 18-fold overshoot in intravesicular alanine concentration in the presence of an inwardly directed Na+ gradient, but not under Na+ equilibrium conditions. A reduced overshoot (10-fold) was seen with an inwardly directed K+ gradient. Further studies revealed a broad cation selectivity, with preference for Na+, which was characteristic of alanine transport but not glucose transport in these membranes. The apparent amino acid specificity of the uptake pathway(s) was similar to that of intact gills and supported the idea of at least four separate pathways for amino acid transport in mussel gill brush border membranes. The apparent Michaelis constant for alanine uptake was approximately 7m, consistent with values forK
t
determined with intact tissue. 相似文献
60.
Thomas K. C. Leung Christine Hall Clinton Monfries Louis Lim 《Journal of neurochemistry》1987,49(1):232-238
Neurone-specific enolase (NSE) and the brain form of creatine phosphokinase (CPK-BB) were previously found to be present in rat synaptosomal plasma membranes (SPM) using two-dimensional gel (2-D gel) and peptide analysis; enzymatic activities of these and of pyruvate kinase (PK), all involved in ATP generation, were shown to be "cryptic" unless the SPM were treated with Triton X-100. We now show that enzymatic activation also occurs when the SPM are treated with trifluoperazine (TFP). TFP activation occurred even when the enzymes were membrane associated, showing that solubilization was not responsible for "unmasking" the enzyme activities. When TFP treatment was performed at alkaline instead of neutral pH, NSE and CPK-BB were released as well as PK, nonneuronal enolase, and aldolase which were identified by 2-D gel and tryptic peptide analysis. Other proteins released included calmodulin, actin, and the 70-kilodalton heat-shock cognate protein. Tubulin, synapsin I, and a 35-kilodalton basic protein were largely unaffected. The latter was identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase on the basis of 2-D gel and peptide analyses and subsequent partial sequencing of a rat brain cDNA coding for the same protein. TFP treatment is thus useful for activating latent enzymes as well as for distinguishing enzymes that have a different disposition on the membrane. 相似文献