全文获取类型
收费全文 | 3459篇 |
免费 | 191篇 |
国内免费 | 1篇 |
出版年
2022年 | 15篇 |
2021年 | 24篇 |
2020年 | 13篇 |
2019年 | 26篇 |
2018年 | 34篇 |
2017年 | 26篇 |
2016年 | 48篇 |
2015年 | 75篇 |
2014年 | 98篇 |
2013年 | 233篇 |
2012年 | 195篇 |
2011年 | 197篇 |
2010年 | 131篇 |
2009年 | 122篇 |
2008年 | 216篇 |
2007年 | 185篇 |
2006年 | 170篇 |
2005年 | 177篇 |
2004年 | 164篇 |
2003年 | 187篇 |
2002年 | 156篇 |
2001年 | 59篇 |
2000年 | 48篇 |
1999年 | 58篇 |
1998年 | 45篇 |
1997年 | 49篇 |
1996年 | 37篇 |
1995年 | 36篇 |
1994年 | 29篇 |
1993年 | 46篇 |
1992年 | 57篇 |
1991年 | 47篇 |
1990年 | 46篇 |
1989年 | 55篇 |
1988年 | 38篇 |
1987年 | 53篇 |
1986年 | 40篇 |
1985年 | 51篇 |
1984年 | 36篇 |
1983年 | 42篇 |
1982年 | 28篇 |
1981年 | 28篇 |
1980年 | 25篇 |
1978年 | 25篇 |
1977年 | 19篇 |
1976年 | 16篇 |
1975年 | 20篇 |
1972年 | 17篇 |
1970年 | 15篇 |
1968年 | 14篇 |
排序方式: 共有3651条查询结果,搜索用时 46 毫秒
11.
Linda Owers Narhi Michael F. Rhode Pamela Hunt Tsutomu Arakawa 《Journal of Protein Chemistry》1989,8(5):669-677
The limited proteolysis of human recombinant TNF- by trypsin yields two stable products resulting from cleavage after Arg6 and Arg44. In solution these two products remain associated together in a trimer with a Stokes' radius slightly greater than the radius of intact TNF- and, therefore, could not be separated from each other under nondenaturing conditions. This limited digest retains at least 20% of the activity of the original TNF- sample, and has a tertiary structure that is similar to that of the native protein by circular dichroism. On the other hand, incorrectly folded, inactive TNF- undergoes extensive digestion following similar treatment with trypsin. These results indicate that the active form of TNF- has a tight core structure which is maintained afterN-terminal cleavage and removal. 相似文献
12.
Fusae Shimizu Mari Ogata Tatsuhiko Yagi Sadao Wakabayashi Hiroshi Matsubara 《Biochimie》1989,71(11-12)
Rubredoxin was purified from Desulfovibrio vulgaris Miyazaki. It was sequenced and some of its properties determined. Rubredoxin is composed of 52 amino acids. It is highly homologous to that from D. vulgaris Hildenborough. Its N-methionyl residue is partially formalated. The millimolar absorption coefficients of the rubredoxin at 489 nm and 280 are 8.1 and 18.5, respectively, and the standard redox potential is +5 mB, which is slightly higher than those of other rubredoxins. Rubredoxin, as well as cytochrome c-553, was reduced with lactate by the action of lactate dehydrogenase of this organism, and the rection was stimulated with 2-methyl-1, 4-naphthoquinone. It is suggested that rubredoxin, in collaboration with membraous quinone, functions as natural electron carrier for cytoplasmic lactate dehydrogenase of this organism, whereas cytochrome c-553 plays the same role for periplasmic lactate dehydrogenase. 相似文献
13.
Yuichi Fujita Yasuhiro Takahashi Takayuki Kohchi Haruo Ozeki Kanji Ohyama Hiroshi Matsubara 《Plant molecular biology》1989,13(5):551-561
The frxC gene, one of the unidentified open reading frames present in liverwort chloroplast DNA, shows significant homology with the nifH genes coding for the Fe protein, a component of the nitrogenase complex (Ohyama et al., 1986, Nature 322: 572–574). A truncated form of the frxC gene was designed to be over-expressed in Escherichia coli and an antibody against this protein was prepared using the purified product as an antigen. This antibody reacted with a protein in the soluble fraction of liverwort chloroplasts, which had an apparent molecular weight of 31 000, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in good agreement with a putative molecular weight of 31945 deduced from the DNA sequence of the frxC gene. In a competitive inhibition experiment, the antigenicity of this protein was indicated to be similar to that of the over-expressed protein in E. coli. Therefore, we concluded that the frxC gene was expressed in liverwort chloroplasts and that its product existed in a soluble form. The molecular weight of the frxC protein was approximately 67 000, as estimated by gel filtration chromatography, indicating that the frxC protein may exist as a dimer of two identical polypeptides analogous to the Fe protein of nitrogenase. The results obtained from affinity chromatography supported the possibility that the frxC protein, which possesses a ATP-binding sequence in its N-terminal region that is conserved among various other ATP-binding proteins, has the ability to bind ATP. 相似文献
14.
Molecular cloning and nucleotide sequence of human pancreatic prechymotrypsinogen cDNA 总被引:4,自引:0,他引:4
N Tomita Y Izumoto A Horii S Doi H Yokouchi M Ogawa T Mori K Matsubara 《Biochemical and biophysical research communications》1989,158(2):569-575
The cDNA clone encoding human prechymotrypsinogen was isolated from a human pancreas cDNA library and its nucleotide sequence was determined. The sequence consists of a 16 bp 5' non-coding region, a 789 bp amino acid coding region and a 60 bp 3' non-coding region. The predicted product consists of 263 amino acids, including 18 amino acids for a signal peptide and 15 amino acids possible for an activation peptide. Southern blot analyses using the cloned cDNA as a probe revealed that human genomic DNA carries at least two genes that are related to chymotrypsinogen. 相似文献
15.
16.
Identification of a novel alpha-amylase by expression of a newly cloned human amy3 cDNA in yeast 总被引:1,自引:0,他引:1
A novel amylase gene (amy3) that differs in nucleotide sequence from salivary amylase gene (amy1) and pancreatic amylase gene (amy2) has been described [Tomita et al., Gene 76 (1989) 11-18], but whether this gene can ever code for an active enzyme has not been shown. We prepared cDNA of this gene from an mRNA obtained from lung carcinoid tissue, and expressed it in Saccharomyces cerevisiae under the control of an acid phosphatase promoter. The product was secreted into culture media, and showed enzymatic activity, demonstrating that this novel alpha-amylase gene (amy3) can code for a functional isozyme. We purified this enzyme, and compared its biological properties with those of salivary and pancreatic human amylases similarly expressed in yeast. We observed that the novel amylase isozyme is more heat-sensitive than others, and that its substrate specificity is different from the other two isozymes. 相似文献
17.
The maximum theanine production by Camellia sinensis cultured cells was achieved by culturing in the modified MS medium containing 2 mg/l indole-3-butyric acid, 0.1 mg/l kinetin, 0 mM NH4NO3 and 39.6 mM KNO3 with 40 mM ethylamine hydrochloride or 20 mM ethylamine hydrochloride and 10 mM L-glutamic acid. Other primary amines, such as methylamine, n-butylamine, 2-hydroxyethylamine, 2cyanoethylamine, aniline, benzylamine and phenylethylamine, were also biotransformed to N5-alkyl-L-glutamine derivatives by C. sinensis cultured cells.Abbreviations IBA
indole-3-butyric acid
- K
kinetin
- MS medium
Murashige and Skoog's basal medium (1962)
- NMR
nuclear magnetic resonance
Part 70 in the series Studies on Plant Tissue Culture. For Part 69 see Furuya et al. (1990). 相似文献
18.
A novel human oncogene, LCA, was assigned to region 2q14----q21 by in situ molecular hybridization. The present regional mapping substantiates the previous assignment that was performed by Southern blot analyses of DNAs from flow-sorted human chromosomes and human-mouse somatic cell hybrids. 相似文献
19.
Masao Eura Tatsuhiko Maehara Tsutomu Ikawa Takeru Ishikawa 《Cancer immunology, immunotherapy : CII》1988,27(2):147-153
Summary Cytotoxicity was induced in lymphocytes (CL) from 10 out of 15 patients by autologous mixed lymphocyte tumor cell culture and further cultivation with recombinant interleukin-2. In cells from 3 of the 10 patients, cytotoxicity was suppressed by more than 50% when autologous peripheral blood mononuclear cells (PBMC) from the patients with large tumors were added to the autologous killing system. The cells responsible for suppressing the cytotoxicity in the effector phase were adherent or nonadherent to plastic depending on the patient examined. The T cell fraction from 1 patient significantly suppressed the cytotoxic activity, and this suppression was seen only in the autologous system. On the other hand, plastic adherent cells but not T cells from PBMC of 2 subjects suppressed the cytotoxic activity of CL. The reason why the main cell population suppressing the CL activity differed among the patients is unclear. However, the findings that the suppression was mostly abrogated following resection of the tumor mass suggested that suppressor cells, either of macrophage lineage or T cells, are induced in patients with a large tumor mass. This speculation is supported by the finding that the PBMC from a patient with tumor recurrence regained the suppressive activity. 相似文献
20.
Electron microscopy of hepatitis B virus core antigen expressing yeast cells by freeze-substitution fixation 总被引:1,自引:0,他引:1
M Yamaguchi T Hirano K Sugahara H Mizokami M Araki K Matsubara 《European journal of cell biology》1988,47(1):138-143
We have used the freeze-substitution fixation technique for electron microscopy of yeast cells that express the hepatitis B virus core antigen (HBcAg) following transformation with the cloned gene. Abundant spherical particles were found within the transformed cells. These particles had a uniform size and shape, measured about 21 nm in diameter, had electron-lucent centers, and consisted of many subunits. They were localized in both the cytoplasm and the nucleus. None of these particles was found in the cells of the parent strain. Comparison of the HBcAg particles isolated from the yeast cells and the particles within the yeast cells demonstrated that the 21-nm particles were in fact ultrastructurally superimposable on HBcAg. Thus, the HBcAg particles within the yeast cells were similar to the HBcAg particles in human liver tissues infected with hepatitis B virus, not only in their size and appearance, but also in their intracellular localization. These results suggest that the yeast cell has the same machinery for synthesis and intracellular translocation of the HBcAg polypeptides as the human cell. 相似文献