全文获取类型
收费全文 | 85276篇 |
免费 | 8383篇 |
国内免费 | 68篇 |
专业分类
93727篇 |
出版年
2022年 | 673篇 |
2021年 | 1246篇 |
2020年 | 903篇 |
2019年 | 1007篇 |
2018年 | 1415篇 |
2017年 | 1209篇 |
2016年 | 1937篇 |
2015年 | 2716篇 |
2014年 | 2768篇 |
2013年 | 3636篇 |
2012年 | 4170篇 |
2011年 | 3950篇 |
2010年 | 2469篇 |
2009年 | 2330篇 |
2008年 | 2937篇 |
2007年 | 2985篇 |
2006年 | 2748篇 |
2005年 | 3030篇 |
2004年 | 2999篇 |
2003年 | 2642篇 |
2002年 | 2295篇 |
2001年 | 3335篇 |
2000年 | 3231篇 |
1999年 | 2704篇 |
1998年 | 1051篇 |
1997年 | 1021篇 |
1996年 | 1086篇 |
1995年 | 888篇 |
1994年 | 899篇 |
1993年 | 867篇 |
1992年 | 2005篇 |
1991年 | 1925篇 |
1990年 | 1841篇 |
1989年 | 1802篇 |
1988年 | 1713篇 |
1987年 | 1598篇 |
1986年 | 1424篇 |
1985年 | 1377篇 |
1984年 | 1173篇 |
1983年 | 955篇 |
1982年 | 706篇 |
1981年 | 685篇 |
1980年 | 647篇 |
1979年 | 1008篇 |
1978年 | 813篇 |
1977年 | 715篇 |
1975年 | 753篇 |
1974年 | 782篇 |
1973年 | 719篇 |
1972年 | 617篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
11.
J G Fukushima O Cascone J A Santomé M J Biscoglio de Jimenez Bonino 《International journal of peptide and protein research》1987,30(3):365-370
Reactivity of histidine residues in equine growth hormone to ethoxyformic anhydride was studied. The existence of two kinetically different sets was demonstrated: one of them including only the slow reacting histidine 169 (k = 0.164 min-1) and the other containing fast reacting histidines 19 and 21 (k = 0.892 min-1). A correlation between the decrease in the capacity to compete with 125I-labeled hormone for rat liver binding sites and the degree of ethoxyformylation of the fast group was found. Circular dichroism studies indicated no significant conformational changes in the protein with all three residues modified. These results fully agree with those obtained for bovine growth hormone which is further evidence supporting the vinculation of histidines 19 and/or 21 with the binding site of these hormones to their specific receptors. 相似文献
12.
13.
To define catalytically essential residues of bacteriophage T7 RNA polymerase, we have generated five mutants of the polymerase, D537N, K631M, Y639F, H811Q and D812N, by site-directed mutagenesis and purified them to homogeneity. The choice of specific amino acids for mutagenesis was based upon photoaffinity-labeling studies with 8-azido-ATP and homology comparisons with the Klenow fragment and other DNA/RNA polymerases. Secondary structural analysis by circular dichroism indicates that the protein folding is intact in these mutants. The mutants D537N and D812N are totally inactive. The mutant K631M has 1% activity, confined to short oligonucleotide synthesis. The mutant H811Q has 25% activity for synthesis of both short and long oligonucleotides. The mutant Y639F retains full enzymatic activity although individual kinetic parameters are somewhat different. Kinetic parameters, (kcat)app and (Km)app for the nucleotides, reveal that the mutation of Lys to Met has a much more drastic effect on (kcat)app than on (Km)app, indicating the involvement of K631 primarily in phosphodiester bond formation. The mutation of His to Gln has effects on both (kcat)app and (Km)app; namely, three- to fivefold reduction in (kcat)app and two- to threefold increase in (Km)app, implying that His811 may be involved in both nucleotide binding and phosphodiester bond formation. The ability of the mutant T7 RNA polymerases to bind template has not been greatly impaired. We have shown that amino acids D537 and D812 are essential, that amino acids K631 and H811 play significant roles in catalysis, and that the active site of T7 RNA polymerase is composed of different regions of the polypeptide chain. Possible roles for these catalytically significant residues in the polymerase mechanism are discussed. 相似文献
14.
15.
16.
17.
Abstract Electrofusion between cells of yeast strains with different genetic markers in isotonic sorbitol solutions leads to high yields of hybrids when 0.1 mM Ca2+ and 0.5 mM Mg2+ salts are aded. On average, 1000–2000 hybrids are obtained when electrofusion is performed (in a helical chamber) compared to a yield of about 40–120 in the absence of these bivalent cations. A further increase in yield can be achieved by the addition of 1 mg/ml albumin, which results in up to 4000 hybrids per experimental run. The entire fusion process leads to very reproducible results in the presence of these substances. 相似文献
18.
19.
20.
The direct binding of porcine pancreatic phospholipase A2 and its zymogen to 1,2-bis(heptanylcarbamoyl)-rac-glycerol 3-sulfate was studied by optical detection of triplet-state magnetic resonance spectroscopy in zero applied magnetic field. The zero-field splittings of the single Trp3 residue undergo significant changes upon binding of phospholipase A2 to lipid. Shifts in zero-field splittings, characterized mainly by a reduction of the E parameter from 1.215 to 1.144 GHz, point to large changes in the Trp3 local environment which accompany the complexing of phospholipase A2 with lipid. This may be attributed to Stark effects caused by the binding of a charged group near Trp3 in the enzyme-lipid complex. The cofactor, Ca2+, which is strongly bound to the enzyme active site, has an influence on the bonding, as reflected by smaller zero-field splitting shifts. A relatively small change in the Trp environment was observed for the interaction of the zymogen with lipid. 相似文献