全文获取类型
收费全文 | 20711篇 |
免费 | 1953篇 |
国内免费 | 21篇 |
专业分类
22685篇 |
出版年
2023年 | 102篇 |
2022年 | 218篇 |
2021年 | 466篇 |
2020年 | 209篇 |
2019年 | 314篇 |
2018年 | 360篇 |
2017年 | 316篇 |
2016年 | 572篇 |
2015年 | 992篇 |
2014年 | 1090篇 |
2013年 | 1285篇 |
2012年 | 1720篇 |
2011年 | 1675篇 |
2010年 | 1107篇 |
2009年 | 889篇 |
2008年 | 1379篇 |
2007年 | 1349篇 |
2006年 | 1226篇 |
2005年 | 1198篇 |
2004年 | 1158篇 |
2003年 | 1079篇 |
2002年 | 1021篇 |
2001年 | 175篇 |
2000年 | 100篇 |
1999年 | 205篇 |
1998年 | 256篇 |
1997年 | 156篇 |
1996年 | 136篇 |
1995年 | 124篇 |
1994年 | 115篇 |
1993年 | 113篇 |
1992年 | 93篇 |
1991年 | 93篇 |
1990年 | 83篇 |
1989年 | 66篇 |
1988年 | 55篇 |
1987年 | 62篇 |
1986年 | 50篇 |
1985年 | 65篇 |
1984年 | 69篇 |
1983年 | 73篇 |
1982年 | 80篇 |
1981年 | 75篇 |
1980年 | 75篇 |
1979年 | 36篇 |
1978年 | 66篇 |
1977年 | 43篇 |
1976年 | 38篇 |
1975年 | 39篇 |
1973年 | 27篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
71.
Eric Giraud Alain Brauman Simon Keleke Bertrand Lelong Maurice Raimbault 《Applied microbiology and biotechnology》1991,36(3):379-383
Summary An amylolytic lactic acid bacterium identified as Lactobacillus plantarum was isolated from cassava roots (Manihot esculenta var. Ngansa) during reting. The amylolytic enzyme synthesized was an extracellular -amylase with an optimum pH of 5.0 and an optimum temperature of 55° C. Cultured on starch, the strain displayed a growth rate of 0.43 h–1, a biomass yield of 0.19 g·g–1 and a lactate yield of 0.81 g·g–1. The growth kinetics were similar on starch and glucose. Sufficient enzyme was synthesized and starch hydrolysis was not a limiting factor for growth. Biosynthesis of the enzyme was observed when the glucose concentration was less than 6.7 g·l–1 and reached up to 4 IU·ml–1 at the end of the fermentation.
Offprint requests to: M. Raimbault 相似文献
72.
Om P. Malhotra Frank Valencic Eric T. Fossel Karl A. Koehler 《Journal of Protein Chemistry》1991,10(1):31-41
Ca2+ titrations of the intrinsic fluorescence of a series of -carboxyglutamic acid (GLA)-deficient bovine prothrombin fragments 1 yield response Hill plot parameters useful for characterization of the metal ion-binding process. 11-, 10-, and 9-GLA fragments 1 exhibitT
m
(the (Ca2+)total concentration at which ln (B/F)=0 in the response Hill plot) values between 0.2 and 0.3 mM. A 22-fold increase inT
m
to 5.4 mM is observed for 8-GLA fragment 1.T
m
decreases to 3.8 mM for the 7- and 6-GLA proteins. The value ofh, about 2.8±0.2 for 11-, 10-, and 9-GLA fragments 1, abruptly decreases to 1.2–1.3 for 8-, 7-, and 6-GLA fragments 1. The observed degree of quenching induced by saturating levels of calcium ions is affected by both changes in the intrinsic fluorescence of the metal ion-free proteins and in the maximum possible degree of quenching in the presence of calcium. The kinetic characteristics of the calcium ion-induced quenching of the intrinsic fluorescence of 6-GLA fragment 1 are identical to those observed in 10-GLA fragment 1, suggesting that the fluorescence quenching observed in the 6- and 10-GLA fragments 1, while different in magnitude, involves similar processes. Observation of an abrupt change in the relative electrophoretic mobilities of 11- to 9-GLA fragments 1 compared to 8- to 6-GLA fragments 1, in the absence or presence of Ca2+, suggests the existence of a major protein conformation change which occurs concomitantly with the noted changes inT
m
andh response Hill plot parameters. Molecular mechanics calculations suggest a structural hypothesis unifying these observations. Central to this model is the presumption of the existence of hydrogen bond-mediated interactions between metal ion-binding sites. 相似文献
73.
74.
Manuela C. Koch Kenneth Ricker Michael Otto Tiemo Grimm Klaus Bender Barbara Zoll Peter S. Harper Frank Lehmann-Horn Reinhardt Rüdel Eric P. Hoffman 《Human genetics》1991,88(1):71-74
Summary Paramyotonia congenita (PC), an autosomal dominant non-progressive muscle disorder, is characterised by cold-induced stiffness followed by muscle weakness. The weakness is caused by a dysfunction of the sodium channel in muscle fibre. Parts of the gene coding for the -subunit of the sodium channel of the adult human skeletal muscle (SCN4A) have been localised on chromosome 17. To investigate the role of this gene in the etiology of PC, a linkage analysis in 17 well-defined families was carried out. The results (z=20.61, =0.001) show that the mutant gene responsible for the disorder is indeed tightly linked to the SCN4A gene. The mutation causing hyperkalemic periodic paralysis (HyperPP) with myotonia has previously been mapped to this gene locus by the same candidate gene approach. Thus, our data suggest that PC and HyperPP are caused by allelic mutations at a single locus on chromosome 17.Dedicated to Professor P. E. Becker on the occasion of his 83rd birthday. 相似文献
75.
Toshiyuki Itaya Eric Fearon Troy Fiesinger Barbara Hunt Bert Vogelstein Philip Frost 《Cancer immunology, immunotherapy : CII》1991,33(4):267-273
Summary The transfection of murine SP1 tumor cells with the hemagglutinin (HA) gene of influenza virus results, after fluorescent-activated cell sorting (FACS), in the selection of high-HA-expressing cell lines called H4A and H4B. Both lines fail to grow in syngeneic animals at doses that result in 100% tumor take of non-transfected tumor cells. Both grow in immunosuppressed mice. SP1 and H4A or H4B cells express few class I major histocompatibility complex (MHC) antigens but do express class II IAk antigens. H4A or H4B cells engender a cytotoxic T lymphocyte (CTL) response but cannot protect against a challenge with SP1 cells. This CTL response is inhibited by anti-CD4 but not anti-CD8 antibodies. Using FACS, we were able to select a population (called H5AK5) with high class-I MHC antigen expression. Like H4A and H4B, H5AK5 cells fail to grow in syngeneic animals but do grow in immunosuppressed mice. However, unlike H4A or H4B, H5AK5 can induce protection against a challenge with 1 × 105 SP1 cells. These studies indicate that the immunogenicity ofHA-transfected SP1 cells may correlate with the cell-surface expression of class II MHC antigens. However, HA-expressing SP1 cells seem able to induce a protective response against a parent SP1 cell challenge only if they also express class I MHC antigens. This view is supported by the observations that SP1 cells expressing murine interleukin-2 do not express class I MHC antigens, fail to grow in syngeneic animals, do grow in immunosuppressed mice but do not protect against a challenge with parental SP1 cells.This work was supported by The Clayton Fund, The Sid W. Richardson Foundation and PHS grants CA 39853 and 41525. Toshiyuki Itaya is a visiting scientist supported by the Smith Education Fund of the Department of Cell Biology. Troy Fiesinger is a summer research investigator sponsored by The University of Texas M. D. Anderson Cancer Center Summer Program for College Students 相似文献
76.
Repression of Meiotic Crossing over by a Centromere (CEN3 ) in SACCHAROMYCES CEREVISIAE 总被引:12,自引:2,他引:10 下载免费PDF全文
The location of the centromere of chromosome III (CEN3) of Saccharomyces cerevisiae has been altered by means of transformation. The frequency of meiotic crossing over in the CEN3-PGK1 and LEU2-CEN3 intervals increases approximately 1.5- and fourfold, respectively, when CEN3 is repositioned at HIS4. The centromere-distal HIS4-LEU2 region experiences a three- to fivefold decrease in the frequency of meiotic exchange when CEN3 is repositioned at HIS4. The inhibition of meiotic crossing over is conferred by a 627-base-pair fragment of CEN3 DNA and is not dependent on the orientation of CEN3 relative to the rest of chromosome III. 相似文献
77.
Distinguishing between Nitrification and Denitrification as Sources of Gaseous Nitrogen Production in Soil 总被引:12,自引:2,他引:10 下载免费PDF全文
The source of N2O produced in soil is often uncertain because denitrification and nitrification can occur simultaneously in the same soil aggregate. A technique which exploits the differential sensitivity of these processes to C2H2 inhibition is proposed for distinguishing among gaseous N losses from soils. Denitrification N2O was estimated from 24-h laboratory incubations in which nitrification was inhibited by 10-Pa C2H2. Nitrification N2O was estimated from the difference between N2O production under no C2H2 and that determined for denitrification. Denitrification N2 was estimated from the difference between N2O production under 10-kPa C2H2 and that under 10 Pa. Laboratory estimates of N2O production were significantly correlated with in situ N2O diffusion measurements made during a 10-month period in two forested watersheds. Nitrous oxide production from nitrification was most important on well-drained sites of a disturbed watershed where ambient NO3− was high. In contrast, denitrification N2O was most important on poorly drained sites near the stream of the same watershed. Distinction between N2O production from nitrification and denitrification was corroborated by correlations between denitrification N2O and water-filled pore space and between nitrification N2O and ambient NO3−. This technique permits qualitative study of environmental parameters that regulate gaseous N losses via denitrification and nitrification. 相似文献
78.
Wounding Inhibits Protein Synthesis yet Stimulates Polysome Formation in Aged, Excised Pea Epicotyls 总被引:2,自引:0,他引:2
Wounding of aged, previously-excised pea epicotyl segments byremoval of the basal 12 mm resulted in a rapid (beginningwithin 15 min) recruitment of monosomes on to polysomes andan even more rapid (maximal between 612 min) inhibitionof protein synthesis in the remaining tissue. This inhibitionof protein synthesis in vivo did not appear to be an artefactcaused by the removal of highly active tissue (e.g., callus,contaminating bacteria), since wounds inflicted at a site distantfrom the region analyzed still elicited the response, and proteinsynthesis in the 12 mm slices (normally discarded) wasinhibited even more strongly than it was in the remaining tissue.The proportion of radioactive methionine in nascent chains (boundto polysomes) increased, while the production of completed polypeptidesdecreased, after wounding. Cycloheximide, a known inhibitorof the ribosome translocation/release process mimicked someof the effects of wounding. We interpret the results to indicatethat the initial effect of wounding is to inhibit translationby inhibiting the ribosome translocation/release process, whereasthe subsequent recovery in protein synthesis is brought aboutpartly by a recovery in ribosome translocation/release and partlyby enhanced initiation.
1 Present address: Harvard-MIT Division of Health Science andTechnology, MIT, Cambridge, Massachusetts 02139, U.S.A.
2 Present address: Institute of Agricultural Environment Control,College of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama790, Japan. (Received May 26, 1986; Accepted August 4, 1986) 相似文献
79.
Rapid Isolation of the 7S-Nerve Growth Factor Complex and Its Subunits from Murine Submaxillary Glands and Saliva 总被引:5,自引:3,他引:2
7S-Nerve growth factor (NGF) and its alpha, beta-NGF, and gamma subunits have been purified from murine submaxillary glands and saliva by a combination of gel filtration on rigid polyvinyl gels, reversed-phase liquid chromatography on short alkyl chain supports (C4 columns), and ion-exchange chromatography on silica-based carboxymethyl columns. This technique is superior to previously used methods in that it is much more rapid and allows the purification of larger quantities of polypeptide from the same amount of starting material. Beta-NGF prepared with this method elicits the outgrowth of fibers of cells of a pheochromocytoma cell line (PC 12) in vitro, indicating that the biological activity is not impaired by the organic solvents and strong acids utilized for its isolation. 相似文献
80.
Michael M. Lipsky Talia R. Sheridan Richard O. Bennett Eric B. May 《In vitro cellular & developmental biology. Plant》1986,22(6):360-362
Summary Comparisons were made of attachment and viability of rainbow trout (Salmo gairdneri) hepatocytes in short-term (2 days), primary culture on plastic, collagen-coated or extracellular matrix (ECM) coated dishes.
Hepatocyte isolation routinely yielded cells with good viability (96%). Cells plated on ECM attached with high efficiency
(93%) in contrast to cells cultured on plastic or collagen (∼20%). The cells plated on ECM flattened out and formed monolayers,
while the cells on plastic and collagen rounded up and formed multi-cell aggregates in suspension. Viability of cells in all
substrates remained high over the 2 day culture period. ECM is the first substrate to support trout-hepatocyte attachment
in primary culture. Differentiated liver function was maintained in cells cultured on ECM as evidence by the induction of
tyrosine aminotransferase by hydrocortisone (200%).
This work was supported in part by research grant R809599010 from the U. S. Environmental Protection Agency.
Editor's Statement This paper reports improved methods for culture of trout liver-derived cells that make in vitro investigations
of fish metabolism, carcinogenesis and chemical toxicity more feasible than previously applied techniques. Recent interest
in fish as models for study and indicators of effects of envionmental and food-related toxins make this work timely, poarticularly
since many of the compounds of interest are primarily metabolized by hepatocytes or act on liver as a major target. David
W. Barnes 相似文献