全文获取类型
收费全文 | 11988篇 |
免费 | 1013篇 |
国内免费 | 426篇 |
出版年
2023年 | 71篇 |
2022年 | 117篇 |
2021年 | 433篇 |
2020年 | 301篇 |
2019年 | 332篇 |
2018年 | 362篇 |
2017年 | 313篇 |
2016年 | 447篇 |
2015年 | 679篇 |
2014年 | 740篇 |
2013年 | 862篇 |
2012年 | 980篇 |
2011年 | 938篇 |
2010年 | 585篇 |
2009年 | 448篇 |
2008年 | 600篇 |
2007年 | 536篇 |
2006年 | 525篇 |
2005年 | 446篇 |
2004年 | 376篇 |
2003年 | 322篇 |
2002年 | 323篇 |
2001年 | 251篇 |
2000年 | 273篇 |
1999年 | 207篇 |
1998年 | 111篇 |
1997年 | 107篇 |
1996年 | 99篇 |
1995年 | 88篇 |
1994年 | 78篇 |
1993年 | 64篇 |
1992年 | 150篇 |
1991年 | 123篇 |
1990年 | 126篇 |
1989年 | 100篇 |
1988年 | 89篇 |
1987年 | 97篇 |
1986年 | 67篇 |
1985年 | 70篇 |
1984年 | 53篇 |
1983年 | 42篇 |
1982年 | 39篇 |
1980年 | 38篇 |
1979年 | 41篇 |
1978年 | 36篇 |
1977年 | 28篇 |
1975年 | 31篇 |
1974年 | 30篇 |
1973年 | 41篇 |
1972年 | 31篇 |
排序方式: 共有10000条查询结果,搜索用时 31 毫秒
71.
Summary The production of somatic embryos from carrot suspension cultures invariably decreases through simple, repeated subculturing.
Extracellular, concentrated compounds extracted from already established embryo culture not only recovered the embryogenic
capability, but also accelerated the embryo production as much as two-fold (up to 1600 embryos/ml) compared with that of a
control culture. Sugars, which were only a small portion of the total concentrate, were excluded as possible causative factors.
It is likely that a protein fraction that is generated directly by competent, embryogenic cultures is important for the restoration
of embryogenic potential. 相似文献
72.
73.
74.
75.
Summary This paper evaluates and extends the novel method of preparing tissue blocks for paraffin sections within 30 to 60 min, that was proposed in early 1985 in a paper by Boonet al. (1986). More than 2 years' additional experience and testing various microwave ovens has led to new protocols reported in this paper. Results are given of testing (i) an especially designed microwave oven for histoprocessing, (ii) microwavable reagents, (iii) processing larger numbers of specimens simultaneously, (iv) handling different types and sizes of tissue. It is concluded that effective temperature control offers substantial advantages. In addition, the possibilities of performing routine diagnostic pathology omitting formalin altogether are sketched. 相似文献
76.
R A Skidgel C D Bennett J W Schilling F L Tan D K Weerasinghe E G Erd?s 《Biochemical and biophysical research communications》1988,154(3):1323-1329
Human plasma carboxypeptidase N was purified to homogeneity and its active and inactive subunits were separated. By introducing a novel technique, both forms of the active subunit (Mr = 55,000 and Mr = 48,000) were isolated. N-terminal sequencing of the active subunit of human carboxypeptidase N revealed significant homology with the N-terminal sequence of bovine carboxypeptidase H (43% identity) and to a lesser extent with carboxypeptidase A (29% identity) or carboxypeptidase B (18% identity). The active subunit of carboxypeptidase N was hydrolyzed with trypsin and 4 of the tryptic peptides were isolated by HPLC and sequenced. The sequences of the four peptides were homologous (39-64% identity) with regions of carboxypeptidase H corresponding to the middle (residues 148-175) and C-terminal portion (residues 321-408). These regions had essentially no homology with carboxypeptidase A or B. These data indicate that carboxypeptidase H and the active subunit of carboxypeptidase N may have diverged from a common ancestral gene. 相似文献
77.
Further characterization and chemical purity assessment of the human erythrocyte glucose transporter preparation 总被引:2,自引:0,他引:2
A L Rampal E K Jung J J Chin M R Deziel H B Pinkofsky C Y Jung 《Biochimica et biophysica acta》1986,859(2):135-142
Chemical and functional purity of the human erythrocyte glucose transporter preparation obtained by DEAE column chromatography after octyl glucoside solubilization was assessed. The cytochalasin B binding capacity of the preparation indicates that the preparation is 60-85% functional glucose transporter. Gel filtration chromatography on TSK 250 column separates this preparation into at least three major peptide fractions, namely, P0, P1 and P2, with apparent Mr of approx. 80 000, 43 000 and 17 000, respectively. When the preparation is photolabelled with [3H]cytochalasin B prior to the separation only P0 and P1 are labelled. Exposure of the preparation to octyl glucoside or to ultraviolet light irradiation results in an increase in P0 in a time-dependent manner with a concomitant and proportional reduction in P1, without affecting P2 appreciably. For individual preparations, relative abundance of P0 and P1 vary widely in a reciprocal fashion, while that of P2 is practically fixed at approx. 10% of the total protein. The specific activity of cytochalasin B binding of each preparation correlates linearly with the relative abundance of P1 of the preparation, which gives a calculated specific binding activity of 22 nmol/mg protein for this fraction. These results indicate that P1 and P0 are native and denatured transporter, respectively, while P2 is contaminating protein impurities. These results demonstrate that the glucose transporter preparation contains approx. 10% of nontransporter protein impurities, with a varying amount (up to 30%) of denatured transporter, and that the transporter free of the chemical impurities and the denatured transporter can be obtained by a gel filtration chromatography of this preparation. 相似文献
78.
R LoBrutto G W Smithers G H Reed W H Orme-Johnson S L Tan J S Leigh 《Biochemistry》1986,25(19):5654-5660
Pulsed electron paramagnetic resonance spectroscopy has been used to detect Mn(II)-ligand superhyperfine couplings in complexes with creatine kinase and in the Mn(II) metalloprotein concanavalin A. Electron spin-echo envelopes from Mn(II), bound in these complexes, are modulated by superhyperfine interactions between Mn(II) and nearby, weakly coupled nuclear spins. The characteristic frequencies of the modulations were obtained by Fourier transformation of the three-pulse, spin-echo envelopes. In transition-state analogue complexes of creatine kinase (enzyme-MnIIADP-anion-creatine), superhyperfine interactions from the directly coordinated nitrogen of the thiocyanate ligand give envelope modulations. The source of the modulations was confirmed by measurements with the 14N and 15N forms of thiocyanate. On the other hand, the nitrogen of coordinated nitrate, which is two bonds removed from the paramagnetic center, does not produce detectable modulations. In spectra for Mn(II) concanavalin A, envelope modulations are detected due to the nitrogen of the coordinated histidine residue. Complexes prepared in 2H2O give strong signals due to weakly coupled 2H. For Mn(II)-doped single crystals of sodium pyrophosphate, signals are observed in the frequency domain spectra that are due to coupling from 31P. Phosphorus signals from the ADP ligand in complexes with creatine kinase show approximately the same coupling constant but have a much broader line width. 相似文献
79.
Epitopes, structural domains, and asymmetry of amino acid residues in SS-B/La nuclear protein 总被引:21,自引:0,他引:21
E K Chan A M Francoeur E M Tan 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(10):3744-3749
SS-B/La is a conserved cellular phosphoprotein of 46 to 48 KD that is the target antigen of autoantibodies in sera of patients with Sjogren's syndrome and systemic lupus erythematosus. SS-B/La is also known to be associated with certain small cellular and viral RNA, including adenovirus VAI and VAII RNA. Two relatively protease-resistant domains (X and Y) were defined in SS-B from HeLa cells by using human autoantibodies as reagents. Domain X, a methionine-containing nonphosphorylated 28 KD polypeptide, was found to be resistant to partial digestion with six different proteases. Similar domains were also found in calf and rabbit SS-B. Domain Y, a 23 KD polypeptide, was detected after limited digestion with S. aureus V8 and trypsin. This domain contained little if any methionine, but all the detectable phosphorylated amino acids. Among 16 anti-SS-B sera tested by immunoblotting, 11 (69%) were reactive with both domains, three (19%) only with domain X, and two (13%) only with domain Y. These results showed that there are at least two distinct antigenic epitopes on the 46 to 48 KD SS-B/La protein, each located on a separate structural domain. The asymmetric distribution of methionine and phosphorylated amino acid residues in SS-B/La show striking similarity to the two reported domains of the adenovirus 72 KD DNA-binding protein, and raises questions concerning functional similarities that await investigation. 相似文献
80.
An auxiliary protein for DNA polymerase-delta from fetal calf thymus 总被引:62,自引:0,他引:62
An auxiliary protein which affects the ability of calf thymus DNA polymerase-delta to utilize template/primers containing long stretches of single-stranded template has been purified to homogeneity from the same tissue. The auxiliary protein coelutes with DNA polymerase-delta on DEAE-cellulose and phenyl-agarose chromatography but is separated from the polymerase on phosphocellulose chromatography. The physical and functional properties of the auxiliary protein strongly resemble those of the beta subunit of Escherichia coli DNA polymerase III holoenzyme. A molecular weight of 75,000 has been calculated from a sedimentation coefficient of 5.0 s and a Stokes radius of 36.5 A. A single band of 37,000 daltons is seen on sodium dodecyl sulfate gel electrophoresis, suggesting that the protein exists as a dimer of identical subunits. The purified protein has no detectable DNA polymerase, primase, ATPase, or nuclease activity. The ability of DNA polymerase-delta to replicate gapped duplex DNA is relatively unaffected by the presence of the auxiliary protein, however, it is required to replicate templates with low primer/template ratios, e.g. poly(dA)/oligo(dT) (20:1), primed M13 DNA, and denatured calf thymus DNA. The auxiliary protein is specific for DNA polymerase-delta; it has no effect on the activity of calf thymus DNA polymerase-alpha or the Klenow fragment of E. coli DNA polymerase I with primed homopolymer templates. Although the auxiliary protein does not bind to either single-stranded or double-stranded DNA, it does increase the binding of DNA polymerase-delta to poly(dA)/oligo(dT), suggesting that the auxiliary protein interacts with the polymerase in the presence of template/primer, stabilizing the polymerase-template/primer complex. 相似文献