全文获取类型
收费全文 | 1555篇 |
免费 | 101篇 |
专业分类
1656篇 |
出版年
2023年 | 8篇 |
2022年 | 7篇 |
2021年 | 30篇 |
2020年 | 7篇 |
2019年 | 13篇 |
2018年 | 19篇 |
2017年 | 14篇 |
2016年 | 26篇 |
2015年 | 39篇 |
2014年 | 48篇 |
2013年 | 61篇 |
2012年 | 87篇 |
2011年 | 84篇 |
2010年 | 41篇 |
2009年 | 41篇 |
2008年 | 71篇 |
2007年 | 88篇 |
2006年 | 91篇 |
2005年 | 91篇 |
2004年 | 77篇 |
2003年 | 67篇 |
2002年 | 66篇 |
2001年 | 56篇 |
2000年 | 67篇 |
1999年 | 50篇 |
1998年 | 17篇 |
1997年 | 14篇 |
1996年 | 18篇 |
1995年 | 13篇 |
1994年 | 13篇 |
1993年 | 10篇 |
1992年 | 24篇 |
1991年 | 26篇 |
1990年 | 19篇 |
1989年 | 29篇 |
1988年 | 33篇 |
1987年 | 30篇 |
1986年 | 25篇 |
1985年 | 15篇 |
1984年 | 15篇 |
1983年 | 7篇 |
1982年 | 18篇 |
1981年 | 10篇 |
1979年 | 16篇 |
1978年 | 4篇 |
1977年 | 6篇 |
1974年 | 4篇 |
1973年 | 7篇 |
1968年 | 5篇 |
1966年 | 5篇 |
排序方式: 共有1656条查询结果,搜索用时 15 毫秒
911.
912.
Dong H Mukaiyama A Tadokoro T Koga Y Takano K Kanaya S 《Journal of molecular biology》2008,378(1):264-272
Ribonuclease HII from hyperthermophile Thermococcus kodakaraensis (Tk-RNase HII) is a kinetically robust monomeric protein. The conformational stability and folding kinetics of Tk-RNase HII were measured for nine mutant proteins in which a buried larger hydrophobic side chain is replaced by a smaller one (Leu/Ile to Ala). The mutant proteins were destabilized by 8.9 to 22.0 kJ mol− 1 as compared with the wild-type protein. The removal of each -CH2- group burial decreased the stability by 5.1 kJ mol− 1 on average in the mutant proteins of Tk-RNase HII examined. This is comparable with the value of 5.3 kJ mol− 1 obtained from experiments for proteins from organisms growing at moderate temperature. We conclude that the hydrophobic residues buried inside protein molecules contribute to the stabilization of hyperthermophilic proteins to a similar extent as proteins at normal temperature. In the folding experiments, the mutant proteins of Tk-RNase HII examined exhibited faster unfolding compared with the wild-type protein. These results indicate that the buried hydrophobic residues strongly contribute to the kinetic robustness of Tk-RNase HII. This is the first report that provides a practical cause of slow unfolding of hyperthermostable proteins. 相似文献
913.
Correlation between the antitumor activity of a polysaccharide schizophyllan and its triple-helical conformation in dilute aqueous solution 总被引:3,自引:0,他引:3
Yanaki T Ito W Tabata K Kojima T Norisuye T Takano N Fujita H 《Biophysical chemistry》1983,17(4):337-342
Eight samples of a polysaccharide schizophyllan ranging in weight-average molecular weight Mw (in water) from 5 x 10(3) to 1.3 x 10(5) were prepared and their antitumor activity (expressed in terms of the tumor inhibition ratio) against Sarcoma 180 ascites, intrinsic viscosities [eta], and gel-filtration chromatograms in aqueous solution were determined. The tumor inhibition ratio was essentially unity for samples with Mw higher than 9 x 10(4), but reduced to zero or even to a negative value when Mw was lower than 10(4). The [eta] data combined with the chromatographic data showed that above Mw approximately 9 x 10(4) the predominant species of schizophyllan in aqueous solution is the previously found rigid triple helix, whereas below Mw approximately 9 x 10(4) both triple helices and single chains coexist in the solution and the fraction of triple helices decreases monotonically to zero as Mw is decreased to 5 x 10(3). From these findings it was concluded that the antitumor potency of schizophyllan in water is related to the amount of triple helices relative to that of single chains. 相似文献
914.
Novel tachykinins such as substance K and neuromedin K have been identified in the mammalian CNS. Bilateral injections of substance K and related peptides as well as of substance P into the dopamine cell body area of the ventral tegmental area caused dose-dependent increases in locomotor activity and rearing, in rats. As this behavior was blocked by the pretreatment with haloperidol (0.1 mg/kg, i.p.), the activation of dopaminergic systems may be involved in the hypermotility induced by the administration of substance K. 相似文献
915.
Tadokoro T You DJ Abe Y Chon H Matsumura H Koga Y Takano K Kanaya S 《Biochemistry》2007,46(25):7460-7468
Ribonuclease (RNase) HI from the psychrotrophic bacterium Shewanella oneidensis MR-1 was overproduced in Escherichia coli, purified, and structurally and biochemically characterized. The amino acid sequence of MR-1 RNase HI is 67% identical to that of E. coli RNase HI. The crystal structure of MR-1 RNase HI determined at 2.0 A resolution was highly similar to that of E. coli RNase HI, except that the number of intramolecular ion pairs and the fraction of polar surface area of MR-1 RNase HI were reduced compared to those of E. coli RNase HI. The enzymatic properties of MR-1 RNase HI were similar to those of E. coli RNase HI. However, MR-1 RNase HI was much less stable than E. coli RNase HI. The stability of MR-1 RNase HI against heat inactivation was lower than that of E. coli RNase HI by 19 degrees C. The conformational stability of MR-1 RNase HI was thermodynamically analyzed by monitoring the CD values at 220 nm. MR-1 RNase HI was less stable than E. coli RNase HI by 22.4 degrees C in Tm and 12.5 kJ/mol in DeltaG(H2O). The thermodynamic stability curve of MR-1 RNase HI was characterized by a downward shift and increased curvature, which results in an increased DeltaCp value, compared to that of E. coli RNase HI. Site-directed mutagenesis studies suggest that the difference in the number of intramolecular ion pairs partly accounts for the difference in stability between MR-1 and E. coli RNases HI. 相似文献
916.
K Takano N Hizuka K Asakawa K Chihara K Sanayama K Shizume 《Endocrinologia japonica》1988,35(5):775-781
Four patients with GH deficiency aged 6-14 years old were treated with hGRF for 6 months. Before the treatment maximal plasma GH responses to 50 micrograms iv hGRF administration were 48.6, 12.1, 24.3 and 13.0 ng/ml, respectively. Human GRF was administered at a dosage of 50-100 micrograms twice a day subcutaneously. In two patients, the growth rate was increased from 2.4 and 2.0 cm/year to 8.2 and 9.8 cm/year, respectively. In the other two patients, the growth rate did not change with hGRF treatment. Plasma somatomedin C levels increased in one patient but did not change in the other three. Antibody to hGRF was observed in two patients during the treatment, but the presence of antibody did not affect the growth rate. 相似文献
917.
Our previous study of six non-Gly to Gly/Ala mutant human lysozymes in a left-handed helical region showed that only one non-Gly residue at a rigid site had unfavorable strain energy as compared with Gly at the same position (Takano et al., Proteins 2001; 44:233-243). To further examine the role of left-handed residues in the conformational stability of a protein, we constructed ten Gly to Ala mutant human lysozymes. Most Gly residues in human lysozyme are located in the left-handed helix region. The thermodynamic parameters for denaturation and crystal structures were determined by differential scanning calorimetry and X-ray analysis, respectively. The difference in denaturation Gibbs energy (DeltaDeltaG) for the ten Gly to Ala mutants ranged from + 1.9 to -7.5 kJ/mol, indicating that the effect of the mutation depends on the environment of the residue. We confirm that Gly in a left-handed region is more favorable at rigid sites than non-Gly, but there is little difference in energetic cost between Gly and non-Gly at flexible sites. The present results indicate that dihedral angles in the backbone conformation and also the flexibility at the position should be considered for analyses of protein stability, and protein structural determination, prediction, and design. 相似文献
918.
Yoshida S Takano Y Moriwake Y Tohno Y Masuda M Tohno S Minami T Yamasaki M Morikawa H Yuri K 《Biological trace element research》2001,80(1):13-22
To elucidate compositional changes of the uterine tube by aging, the authors studied age-related changes of elements in human uterine tubes by inductively coupled plasma-atomic emission spectrometry. The uterine tubes were resected postmortem or surgically removed from patients with uterine myoma. It was found that the contents of calcium and magnesium increased progressively with aging in uterine tubes, whereas the contents of phosphorus and iron decreased gradually with aging. The sulfur content of uterine tubes remained constant and independent of aging. Regarding relationships between elements, significant relationships were found between calcium and magnesium contents, between phosphorus and iron contents, between phosphorus and sulfur contents, and between phosphorus and sodium contents in human uterine tubes. 相似文献
919.
Takano K Scholtz JM Sacchettini JC Pace CN 《The Journal of biological chemistry》2003,278(34):31790-31795
We previously suggested that proteins gain more stability from the burial and hydrogen bonding of polar groups than from the burial of nonpolar groups (Pace, C. N. (2001) Biochemistry 40, 310-313). To study this further, we prepared eight Thr-to-Val mutants of RNase Sa, four in which the Thr side chain is hydrogen-bonded and four in which it is not. We measured the stability of these mutants by analyzing their thermal denaturation curves. The four hydrogen-bonded Thr side chains contribute 1.3 +/- 0.9 kcal/mol to the stability; those that are not still contribute 0.4 +/- 0.9 kcal/mol to the stability. For 40 Thr-to-Val mutants of 11 proteins, the average decrease in stability is 1.0 +/- 1.0 kcal/mol when the Thr side chain is hydrogen-bonded and 0.0 +/- 0.5 kcal/mol when it is not. This is clear evidence that hydrogen bonds contribute favorably to protein stability. In addition, we prepared four Val-to-Thr mutants of RNase Sa, measured their stability, and determined their crystal structures. In all cases, the mutants are less stable than the wild-type protein, with the decreases in stability ranging from 0.5 to 4.4 kcal/mol. For 41 Val-to-Thr mutants of 11 proteins, the average decrease in stability is 1.8 +/- 1.3 kcal/mol and is unfavorable for 40 of 41 mutants. This shows that placing an [bond]OH group at a site designed for a [bond]CH3 group is very unfavorable. So, [bond]OH groups can contribute favorably to protein stability, even if they are not hydrogen-bonded, if the site was selected for an [bond]OH group, but they will make an unfavorable contribution to stability, even if they are hydrogen-bonded, when they are placed at a site selected for a [bond]CH3 group. The contribution that polar groups make to protein stability depends strongly on their environment. 相似文献
920.
Effect of pH value of freeze-drying solution on the chromosome integrity and developmental ability of mouse spermatozoa 总被引:8,自引:0,他引:8
The nuclei of freeze-dried mouse spermatozoa are able to retain their chromosome integrity and developmental potential. To optimize the conditions of freeze-drying, we examined whether pH values of the freeze-drying solution affect the chromosome integrity and developmental potential of sperm nuclei. The sperm freeze-drying solution we used contained a high concentration (50 mM) of calcium-chelating EGTA. Sperm chromosomes were examined at the metaphase of the first mitosis after injection of freeze-dried spermatozoa into matured oocytes. The developmental potential of sperm nuclei was assessed by examining the development of fetuses in midgestation. The results showed that both sperm chromosomes and sperm developmental potential are maintained better when the freeze-drying solution was slightly alkaline (pH 8.0) rather than near neutral or acidic (pH 7.4-6.0). The data indicated that the chromosome integrity and developmental ability of mouse spermatozoa are affected by the pH value of freeze-drying solution. 相似文献