全文获取类型
收费全文 | 18440篇 |
免费 | 1648篇 |
国内免费 | 10篇 |
专业分类
20098篇 |
出版年
2022年 | 131篇 |
2021年 | 208篇 |
2020年 | 146篇 |
2019年 | 195篇 |
2018年 | 241篇 |
2017年 | 238篇 |
2016年 | 341篇 |
2015年 | 701篇 |
2014年 | 764篇 |
2013年 | 966篇 |
2012年 | 1174篇 |
2011年 | 1224篇 |
2010年 | 804篇 |
2009年 | 756篇 |
2008年 | 1030篇 |
2007年 | 1113篇 |
2006年 | 1045篇 |
2005年 | 1053篇 |
2004年 | 1027篇 |
2003年 | 853篇 |
2002年 | 854篇 |
2001年 | 269篇 |
2000年 | 219篇 |
1999年 | 245篇 |
1998年 | 275篇 |
1997年 | 165篇 |
1996年 | 137篇 |
1995年 | 126篇 |
1994年 | 142篇 |
1993年 | 130篇 |
1992年 | 158篇 |
1991年 | 142篇 |
1990年 | 123篇 |
1989年 | 153篇 |
1988年 | 114篇 |
1987年 | 137篇 |
1986年 | 113篇 |
1985年 | 147篇 |
1984年 | 128篇 |
1983年 | 120篇 |
1982年 | 136篇 |
1981年 | 121篇 |
1980年 | 100篇 |
1979年 | 94篇 |
1978年 | 83篇 |
1977年 | 97篇 |
1976年 | 74篇 |
1974年 | 94篇 |
1973年 | 83篇 |
1972年 | 67篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
91.
Secondary metabolite biosynthesis in cultured cells of Catharanthus roseus (L.) G. Don immobilized by adhesion to glass fibres 总被引:1,自引:0,他引:1
Summary Suspension-cultured cells of Catharanthus roseus (L.) G. Don were immobilized on glass fibre mats and cultivated in shake flasks. The highly-aggregated immobilized cells exhibited a slower growth rate and accumulated reduced levels of tryptamine and indole alkaloids, represented by catharanthine and ajmalicine, in comparison to cells in suspension. The increased total protein synthesis in immobilized cells suggests a diversion of the primary metabolic flux toward protein biosynthetic pathways and away from other growth processes. In vitro assays for the specific activity of tryptophan decarboxylase (TDC) and tryptophan synthase (TS) suggest that the decreased accumulation of tryptamine in immobilized cells was due to reduced tryptophan biosynthesis. The specific activity of TDC was similar in immobilized and suspension-cultured cells. However, the expression of TS activity in immobilized cells was reduced to less than 25% of the maximum level in suspension-cultured cells. The reduced availability of a free tryptophan pool in immobilized cells is consistent with the reduced TS activity. Reduced tryptamine accumulation, however, was not responsible for the decreased accumulation of indole alkaloids in immobilized cells. Indole alkaloid accumulation increased to a similar level in immobilized and suspension-cultured cells only after the addition of exogenous secolaganin to the culture medium. The addition of tryptophan resulted in increased accumulation of tryptamine, but had no effect on indole alkaloid levels. Reduced biosynthesis of secologanin, the monoterpenoid precursor to indole alkaloids, in immobilized cells is suggested. Immobilization does not appear to alter the activity of indole alkaloid biosynthetic enzymes in our system beyond, and including, strictosidine synthase.
Offprint requests to: P. J. Facchini 相似文献
92.
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
93.
Determination of the origin of nondisjunction in a 49,XXXXY male using hypervariable dinucleotide repeat sequences 总被引:1,自引:1,他引:0
Summary We present a patient with a 49,XXXXY chromosome constitution in whom the origin of the extra X chromosomes was determined by analysis of five polymorphic CA (or GT) dinucleotide repeat sequences. This class of DNA marker has recently been demonstrated to be hypervariable with heterozygosity values up to 80%. By polymerase chain reaction (PCR) analysis of the dinucleotide repeat length polymorphisms, we have shown that all four X chromosomes were of maternal origin. 相似文献
94.
Lymphocyte activation and serine-esterase induction following recombinant interleukin-2 infusion for lymphomas and acute leukaemias 总被引:3,自引:0,他引:3
Seah H. Lim Colin Worman Andrew Jewell Chris Tsakona Frank J. Giles Anthony Goldstone 《Cancer immunology, immunotherapy : CII》1991,33(2):133-137
Summary C57BL mice inoculated with radiation leukemia virus (RadLV) develop preleukemic cells long before the onset of leukemia. These cells are potentially immunogenic but fail to elicit an immune response in the host because of the appearance of virus-specific suppressor T cells. We have studied the effect of polysaccharide K (PSK) on the generation of RadLV-specific cell-mediated immune responses in vitro. Long-term exposure to PSK in culture potentiated the ability of immunized T cells to respond to a RadLV-induced lymphoma. It also abrogated the suppressive activity of suppressor T cells and simultaneously boosted the ability of reactive T cells to respond. The dual immunostimulating activity of PSK resulted in the generation of T cytotoxic lymphocytes that could lyse lymphoma cells in vitro. The results suggest that PSK could be used as a prophylactic immune response modifier in preleukemia. 相似文献
95.
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
96.
Jessica E. Hoogendijk Gerard W. Hensels Ina Zorn Linda Valentijn Emiel A. M. Janssen Marianne de Visser David F. Barker Bram W. Ongerboer de Visser Frank Baas Pieter A. Bolhuis 《Human genetics》1991,88(2):215-218
Summary Recently, it has been shown that Charcot-Marie-Tooth disease type 1a (CMT1a) is linked with a duplication of a DNA segment that is detected by probe VAW409R3, and that is located on chromosome 17p11.2. Here, we show that this duplication also contains VAW412R3a, but not A10-41 and EW503. Accounting for the duplication in recombination analysis, we found recombinants between CMT1a and EW301 and EW502, but not with A10-41, VAW409R3, and VAW412R3. Using pulsed-field gel electrophoresis analysis, we estimated the minimal size of the duplicated region in CMT1a patients to be 1100 kb. 相似文献
97.
Expression of the chemically synthesized gene for ribonuclease T1 in Escherichia coli using a secretion cloning vector 总被引:3,自引:0,他引:3
R Quaas Y McKeown P Stanssens R Frank H Bl?cker U Hahn 《European journal of biochemistry》1988,173(3):617-622
The gene for ribonuclease T1 from Aspergillus oryzae has been chemically synthesized using the segmental support technique. An Escherichia coli clone producing the ribonuclease at high levels was constructed by linking the gene downstream to the region coding for the signal peptide of the OmpA protein (a major outer membrane protein of E. coli), using the secretion cloning vector pIN-III-ompA2. This strategy was employed in order to circumvent a possible toxic effect of the gene product on the host cell. Active ribonuclease containing four additional amino acids at the N-terminus could be isolated from the periplasmic fraction of the host. The final yield after purification was 20 mg enzyme/l liquid culture. With respect to immunological, catalytic and specific behaviour, no qualitative differences could be detected between the enzyme from the over-producing E. coli strain and ribonuclease T1 isolated from A. oryzae. 相似文献
98.
Z L Hegedus H A Frank T I Steinman M D Altschule U Nayak 《Archives internationales de physiologie et de biochimie》1988,96(5):211-221
The fluorescence excitation and emission spectra observed in plasma from patients with chronic renal failure were reproduced by the generation of soluble lipofuscins in normal plasma samples by incubation with mixtures of L-dopa, dopamine, L-norepinephrine, L-epinephrine, 3-hydroxy-DL-kynurenine and 3-hydroxy-anthranilic acid for 24 h at 37 degrees C. Relative fluorescence intensity measurements consistently showed elevated plasma levels of the soluble lipofuscins in chronic renal failure: the means (n = 27) were 73.9 +/- 33.4 (SD) and 71.1 +/- 14.8 at emissions 413 nm and 445 nm respectively, in contrast to those of normal plasma samples (n = 11), 18.2 +/- 5.3 and 23.1 +/- 5.6. The maximum or shoulder at approximately 413 nm represents soluble lipofuscin that can be generated from 3-hydroxyanthranilic acid and the maximum or shoulder at approximately 445 nm represents soluble lipofuscins derived from the precursors listed above and probably from other related precursors. Gravimetric measurements also showed elevated levels of melanins in the plasma samples of patients with chronic renal failure: 2.72 +/- 0.38 mg/ml (n = 16), as compared to normal values: 1.70 +/- 0.10 mg/ml (n = 6). In individual patients haemodialysis reduced the fluorescence intensities to a range of 65-99% and the melanin levels to a range of 86-99% of the pre-dialysis values. 相似文献
99.
Immunochemical detection of the alpha-subunit of the G-protein, GZ, in membranes and cytosols of mammalian cells 总被引:1,自引:0,他引:1
K Spicher K D Hinsch H Gausepohl R Frank W Rosenthal G Schultz 《Biochemical and biophysical research communications》1988,157(3):883-890
An antiserum (AS 98) was raised against a synthetic peptide deduced from published cDNA sequences of the alpha-subunit of the putative G-protein, GZ (Fong et al. Proc. Natl. Acad. Sci. USA 85, 3066-3070, 1988; Matsuoka et al. Proc. Natl. Acad. Sci. USA 85, 5384-5388, 1988). In membrane and cytosol preparations of many but not all tested mammalian tissues, AS 98 predominantly recognized two proteins of 40 and 43 kDa Mr. Whereas high levels of a 40 kDa GZ alpha-subunit were found in rat liver membranes and in brain cytosol, AS 98 failed to detect the alpha-subunit of GZ in brain membranes. 相似文献
100.
Kenneth D. Vernick Frank H. Collins Douglas C. Seeley Robert W. Gwadz Louis H. Miller 《Biochemical genetics》1988,26(5-6):367-379
The main polymorphic system of esterase isoenzymes in adults of the G3 laboratory strain ofAnopheles gambiae consists of two to five major bands of activity per individual. The bands are designated 5S, 5F, 13, 14, and 15. In genetic
crosses, the genes which coded for the bands assorted as three codominant alleles, Est A, Est B, and Est C, at a single autosomal
locus. Homozygotes for the Est C allele were significantly underrepresented among backcross progeny. The developmental pattern
of esterase expression was examined. Esterase gene expression in embryos was first detectable between 2 and 12 hr after oviposition.
The initiation or termination of expression of some of the bands corresponded to boundaries between developmental stages.
Most of the esterase fractions were not specifically localized within the tissues tested, with the exception of a series of
bands which were restricted largely to adult male testes. 相似文献