全文获取类型
收费全文 | 6546篇 |
免费 | 420篇 |
国内免费 | 2篇 |
出版年
2022年 | 34篇 |
2021年 | 79篇 |
2020年 | 50篇 |
2019年 | 59篇 |
2018年 | 99篇 |
2017年 | 89篇 |
2016年 | 137篇 |
2015年 | 207篇 |
2014年 | 218篇 |
2013年 | 438篇 |
2012年 | 403篇 |
2011年 | 410篇 |
2010年 | 274篇 |
2009年 | 252篇 |
2008年 | 397篇 |
2007年 | 434篇 |
2006年 | 379篇 |
2005年 | 348篇 |
2004年 | 370篇 |
2003年 | 344篇 |
2002年 | 301篇 |
2001年 | 87篇 |
2000年 | 93篇 |
1999年 | 107篇 |
1998年 | 80篇 |
1997年 | 79篇 |
1996年 | 55篇 |
1995年 | 69篇 |
1994年 | 53篇 |
1993年 | 58篇 |
1992年 | 54篇 |
1991年 | 77篇 |
1990年 | 74篇 |
1989年 | 60篇 |
1988年 | 57篇 |
1987年 | 58篇 |
1986年 | 47篇 |
1985年 | 68篇 |
1984年 | 45篇 |
1983年 | 45篇 |
1982年 | 43篇 |
1981年 | 46篇 |
1980年 | 25篇 |
1979年 | 26篇 |
1978年 | 29篇 |
1977年 | 24篇 |
1976年 | 27篇 |
1975年 | 22篇 |
1974年 | 23篇 |
1973年 | 18篇 |
排序方式: 共有6968条查询结果,搜索用时 15 毫秒
81.
K Furukawa M Tagaya M Inouye J Preiss T Fukui 《The Journal of biological chemistry》1990,265(4):2086-2090
Glycogen synthases from Escherichia coli and mammalian muscle differ in many respects including regulation, sugar nucleotide specificity, and primary sequence. To compare the structure of the active sites in these enzymes, the affinity-labeling study of the E. coli enzyme was carried out using adenosine diphosphopyridoxal as the reagent. The E. coli enzyme was inactivated in a time- and dose-dependent manner when incubated with the reagent followed by sodium borohydride reduction. The inactivation was markedly protected by ADP-glucose and ADP, suggesting that the reagent was bound to the substrate-binding site. The stoichiometry of the bound reagent to the enzyme was approximately 1:1. Sequence analysis of the labeled peptide isolated from a proteolytic digest of the modified protein revealed that Lys15 is labeled. Based on the geometry of the reagent, the epsilon-amino group of this residue might be located close to the pyrophosphate moiety of ADP-glucose bound to the E. coli enzyme, like that of Lys38 in the rabbit muscle enzyme, which is labeled by uridine diphosphopyridoxal (Tagaya, M., Nakano, K., and Fukui, T. (1986) J. Biol. Chem. 260, 6670-6676; Mahrenholz, A. M., Wang, Y., and Roach, P. J. (1988) J. Biol. Chem. 263, 10561-10567). The importance of the conserved sequence of Lys-X-Gly-Gly is discussed in connection with the glycine-rich region found in many nucleotide-binding proteins. 相似文献
82.
Membrane Viscosity Correlates with α1 -Adrenergic Signal Transduction of the Aged Rat Cerebral Cortex
Atsushi Miyamoto Tsunehisa Araiso Tomiyasu Koyama Hideyo Ohshika 《Journal of neurochemistry》1990,55(1):70-75
We investigated, using adult (2-month-old) and senescent (12- and 24-month-old) rats, the effects of aging on the relationship between the alpha 1-adrenergic coupling system and the membrane viscosity of the cerebral cortex. There was no age-related difference in the KD values of [3H]prazosin binding on the membranes. The Bmax values of [3H]prazosin binding were reduced with advanced age. Norepinephrine-induced formation of 3H-labeled inositol phosphates (3H-IPs) in the slices increased with advanced age. The EC50 values for norepinephrine to stimulate the formation of 3H-IPs at advanced age were lower than that at adult age. The cholesterol content in membranes increased with advanced age. No changes in the phospholipid content in membranes were observed with advanced age. Concomitantly, an increase of the molar ratio of cholesterol to phospholipids was observed with advanced age. The membrane viscosity as measured by 1,6-diphenyl-1,3,5-hexatriene increased with advanced age. These results indicate that the altered cholesterol content and/or viscosity in cortical membranes of the aged rat may account for the loss of alpha 1-adrenergic receptor density and/or compensatory changes in the receptor-phospholipase C coupling system. 相似文献
83.
Isolation of Glutamyltaurine from Bovine Brains and Proof of Its γ-Linkage by the B/E Linked Scan SIMS Technique 总被引:1,自引:1,他引:0
Ko Nakamura Kunihiko Higashiura Naoharu Nishimura Atsushi Yamamoto Ikuo Tooyama Hiroshi Kimura Kazuharu Ienaga 《Journal of neurochemistry》1990,55(3):1064-1066
We isolated a glutamyltaurine from bovine brains and determined its structure as gamma-glutamyltaurine (gamma-Glu-Tau; glutaurine) by use of a new mass spectrometric technique [B/E linked scan sputtered ion mass spectrometry (SIMS)], which we have recently shown to be useful for distinguishing the gamma- from the alpha-isomer of glutamyl-dipeptides. Neither the alpha-isomer of glutamyltaurine nor any aspartyltaurines could be detected in bovine brain. 相似文献
84.
85.
Reduction of tetrazolium salts by sulfate-reducing bacteria 总被引:2,自引:0,他引:2
Abstract The reduction of tetrazolium salts by the sulfate-reducing bacteria, Desulfovibrio desulfuricans and Desulfotomaculum orientis , was examined. D. desulfuricans and D. orientis reduced triphenyltetrazolium chloride (TTC) and 2-( p -iodophenyl)-3-( p -nitrophenyl)-5-phenyltetrazolium chloride (INT) forming intracellular formazan deposits. The reduction rate of INT was higher than that of TTC. INT reduction was not inhibited by the addition of sulfate or molybdate, and sulfate uptake was inhibited by the addition of both INT and molybdate. The ratio of intracellular formazan forming cells to acridine orange direct counts in both strains decreased with culture age and starvation time. 相似文献
86.
Production and characterization of recombinant human neutrophil chemotactic factor 总被引:15,自引:0,他引:15
R Furuta J Yamagishi H Kotani F Sakamoto T Fukui Y Matsui Y Sohmura M Yamada T Yoshimura C G Larsen 《Journal of biochemistry》1989,106(3):436-441
A putative mature human neutrophil chemotactic factor (NCF) corresponding to the C-terminal 72 amino acids of its precursor was directly produced in Escherichia coli by recombinant DNA technology. Human NCF was present in both the soluble and insoluble protein fractions of the homogenate of host cells, and it was partially purified as a water-soluble polypeptide from both fractions, separately. The partially purified NCF preparation was highly purified to an endotoxin-free homogeneous polypeptide by means of CM-Sepharose CL-6B column chromatography and gel filtration on Toyopearl HW-55. No difference between the human NCF preparations purified from both starting materials could be found concerning purity, primary structure, solubility, molecular weight, and chemotactic activity for human neutrophils. The amino acid sequence of recombinant human NCF was identical to the sequence deduced from the cDNA sequence. A methionine residue due to the translation initiation codon was removed. Recombinant human NCF was found to be biologically active and to exhibit chemotactic activity for human neutrophils in vitro and cause a neutrophil infiltration in vivo in mice. 相似文献
87.
M Tagaya T Yagami T Noumi M Futai F Kishi A Nakazawa T Fukui 《The Journal of biological chemistry》1989,264(2):990-994
Proline 17 in the glycine-rich region of adenylate kinase was replaced by Gly (the Gly-mutant) or Val (the Val-mutant) by site-directed mutagenesis. The mutant enzymes were purified to homogeneous states on sodium dodecyl sulfate-gel electrophoresis after solubilization of the proteins from the pellets of cell lysates of Escherichia coli. The apparent Km values of the Gly- and the Val-mutants for AMP increased approximately 7- and 24-fold, respectively, as compared with that of the wild-type enzyme. The apparent Km values for ATP also increased 7- and 42-fold in the Gly- and Val-mutants, respectively. In contrast, Vmax values of both mutant enzymes were comparable to that of the wild-type enzyme. These results suggest that Pro-17 plays an important role for the binding of substrates, but not for catalytic efficiency, although it does not directly interact with substrates. Adenosine diphosphopyridoxal, which specifically modifies Lys-21 in adenylate kinase (Tagaya, M., Yagami, T., and Fukui, T. (1987) J. Biol. Chem. 262, 8257-8261), inactivated the wild-type and mutant enzymes at almost the same rates. Interestingly, both mutant enzymes showed higher specificities for adenine nucleotides than the wild-type enzyme. Both mutant enzymes were less resistant than the wild-type enzyme against inactivation at elevated temperatures or by treatment with trypsin. It would appear that most of the properties of the mutant enzymes may be explained on the basis of a need for conformational flexibility of the loop which includes Pro-17 for substrate binding. 相似文献
88.
Naoto Ohnishi Hiroaki Kodania Satoshi Ando Atsushi Komamine 《Physiologia plantarum》1990,80(1):95-101
The effects of inhibition of the synthesis of protein, mRNA or rRNA on the progression of the cell cycle have been analyzed in cultures of Catharanthus roseus in which cells were induced to divide in synchrony by the double phosphate starvation method. The partial inhibition of protein synthesis at the G1 phase by anisoniycio or cycloheximide caused the arrest of cells in the G1 phase or delayed the entry of cells into the S phase. When protein synthesis was partially inhibited at the S phase, cell division occurred to about the same extent as in the control. When asynchronously dividing cells were treated with cycloheximide, cells accumulated in the G1 phase, as shown by flow-cytometric analysis. The partial inhibition of mRNA synthesis by α-amanitin at the G1 phase caused the arrest of cells in the G1 phase, although partial inhibition of mRNA synthesis at the S phase had little effect on cell division. In the case of inhibition of synthesis of rRNA by actinomycin D at the G1 phase, initiation of DNA synthesis was observed, but no subsequent DNA synthesis or the division of cells occurred. However, the addition of actinomycin D during the S phase had no effect on cell division. These results suggest that specific protein(s), required for the progression of the cell cycle, are synthesized in the G1 phase, and that the mRNA(s) that encode these proteins are also synthesized at the G1 phase. 相似文献
89.
Sera (fetal calf serum: FCS; and oestrous cow serum: ECS), hormones (2.5 FSH micrograms/ml + 5 micrograms LH/ml + 1 microgram oestradiol/ml) and granulosa cells (5 x 10(6)/ml) were added to culture medium to determine the frequencies of in-vitro maturation, fertilization, cleavage (2- to 8-cell) and development into blastocysts of bovine follicular oocytes. The maturation rates after 24 h in culture were not significantly different among the three factors tested (56-72%). The fertilization rates were significantly affected by serum type and the addition of granulosa cells. FCS gave significantly higher rates of fertilization (57-71%) than did ECS (34-52%), but the proportions of polyspermic fertilization were significantly higher in the former (8-19%) than in the latter (2-3%). The addition of hormones did not affect fertilization, cleavage and development. Neither type of serum affected cleavage and development. The highest rates of blastocyst formation were obtained when granulosa cells alone were added (FCS, 17%; ECS, 16%). The cell numbers of the blastocysts obtained were 100-150, similar to those of blastocysts developed in vivo. Transfer of 6 blastocysts to 3 cows resulted in 1 pregnancy. The present results indicate that the co-culture with granulosa cells is the most important factor for in-vitro fertilization to development into blastocysts of bovine oocytes matured in vitro. 相似文献
90.