全文获取类型
收费全文 | 4036篇 |
免费 | 367篇 |
国内免费 | 12篇 |
出版年
2021年 | 74篇 |
2020年 | 24篇 |
2019年 | 21篇 |
2018年 | 57篇 |
2017年 | 30篇 |
2016年 | 138篇 |
2015年 | 277篇 |
2014年 | 211篇 |
2013年 | 227篇 |
2012年 | 370篇 |
2011年 | 336篇 |
2010年 | 174篇 |
2009年 | 135篇 |
2008年 | 204篇 |
2007年 | 187篇 |
2006年 | 161篇 |
2005年 | 172篇 |
2004年 | 153篇 |
2003年 | 166篇 |
2002年 | 103篇 |
2001年 | 105篇 |
2000年 | 72篇 |
1999年 | 71篇 |
1998年 | 42篇 |
1997年 | 21篇 |
1996年 | 36篇 |
1995年 | 32篇 |
1994年 | 34篇 |
1993年 | 36篇 |
1992年 | 38篇 |
1991年 | 34篇 |
1990年 | 50篇 |
1989年 | 25篇 |
1988年 | 34篇 |
1987年 | 35篇 |
1986年 | 21篇 |
1985年 | 29篇 |
1984年 | 11篇 |
1983年 | 17篇 |
1981年 | 11篇 |
1980年 | 15篇 |
1979年 | 15篇 |
1978年 | 12篇 |
1975年 | 11篇 |
1973年 | 12篇 |
1972年 | 8篇 |
1970年 | 10篇 |
1969年 | 9篇 |
1968年 | 9篇 |
1927年 | 9篇 |
排序方式: 共有4415条查询结果,搜索用时 15 毫秒
51.
Barnett YA Eger K Eriksson S Folkers G Hansen PE Hofbauer R Komitowsky D Milon A Munch-Petersen B;European Thymidine Kinase Study Group 《Biotechnology advances》1994,12(4):663-668
A precondition for the chemotherapeutic treatment of a variety of virally-induced human diseases and malignant conditions is a highly selective interaction of the drug molecule to be used with it's biological target. To ensure the development of novel, effective drugs, it is essential that the biological target is well characterised with regard to it's structure and activity. Such characterisation relies upon adequate amounts of pure target being available. One of the most important enzymatic importers for antimetabolites is the enzyme thymidine kinase. In this article an in vitro protein expression system is described which facilitates the production of milligram amounts of pure and biologically active thymidine kinase, from a number of important biological sources. Results have shown that the in vitro produced enzyme has the exact biochemical propeties of the in vivo enzyme. Thus the in vitro protein expression system is an ideal vechicle to facilitate an in depth investigation of the enzyme's biological properties. 相似文献
52.
Adherent recombinant BHK cells were cultivated at temperatures between 30 and 37°C. Batch and repeated-batch-cultivations in a 2-litre bioreactor showed a significant influence on metabolism and cell growth. The low-temperature-cultivations showed a lower growth rate and a lower glucose consumption rate and, therefore, less lactate production. On the other hand, the maximum cell density and productivity seemed not to be affected by the temperature reduction. 相似文献
53.
WIP1, a wound-inducible gene from maize with homology to Bowman-Birk proteinase inhibitors 总被引:6,自引:0,他引:6
We have cloned and sequenced a wound-inducible cDNA clone designated WIP1 (for wound-induced protein) from maize coleoptiles. It was isolated by differential screening of a cDNA library prepared from excised maize coleoptile segments. The deduced amino acid sequence predicts a secretory, cysteine-rich protein of 102 residues with a calculated molecular mass of 11 kDa and a typical N-terminal signal sequence. The protein has about 30% identity with various Bowman-Birk type proteinase inhibitors. Most interestingly, it is novel in that it is double-headed with exclusive specificity for chymotrypsin. WIP1 is strongly wound-induced in contrast to other members of the Bowman-Birk proteinase inhibitor family, which occur in seeds and are regulated during development. The response is fast, similar to defenceinduced genes, and measurable as early as 30 min after wounding. Induction can also be evoked in the intact coleoptiles and the signal is systemically transmitted in the coleoptile to adjacent regions of the wounded area. Isolation and analysis of the corresponding genomic clone reveals that WIP1 contains an intron of 90 nucleotides. 相似文献
54.
55.
Jaromir Konecny Michael Eckert Michael Schöniger G. Ludwig Hofacker 《Journal of molecular evolution》1993,36(5):407-416
Summary We lay new foundations to the hypothesis that the genetic code is adapted to evolutionary retention of information in the
antisense strands of natural DNA/RNA sequences. In particular, we show that the genetic code exhibits, beyond the neutral
replacement patterns of amino acid substitutions, optimal properties by favoring simultaneous evolution of proteins encoded
in DNA/RNA sense-antisense strands. This is borne out in the sense-antisense transformations of the codons of every amino
acid which target amino acids physicochemically similar to each other. Moreover, silent mutations in the sense strand generate
conservative ones in its antisense counterpart and vice versa. Coevolution of proteins coded by complementary strands is shown
to be a definite possibility, a result which does not depend on any physical interaction between the coevolving proteins.
Likewise, the degree to which the present genetic code is dedicated to evolutionary sense-antisense tolerance is demonstrated
by comparison with many randomized codes. Double-strand coding is quantified from an information-theoretical point of view. 相似文献
56.
Heilmeyer Ludwig M. G. Gerschinski Andrea M. Meyer Helmut E. Jennissen Herbert P. 《Molecular and cellular biochemistry》1993,127(1):19-30
Holophosphorylase kinase was digested with Glu-C specific protease; from the peptide mixture calmodulin binding peptides were isolated by affinity chromatography and identified by N-terminal sequence analysis. Two peptides originating from the subunit, having a high tendency to form a positively charged amphiphilic helix and containing tryptophane, were synthesized. Additionally, a homologous region of the subunit and a peptide from the subunit present in a region deleted in the isoform were also selected for synthesis. Binding stoichiometry and affinity were determined by following the enhancement in tryptophane fluorescence occurring upon 1:1 complex formation between these peptides and calmodulin. Finally, Ca2+ binding to calmodulin in presence of peptides was measured. By this way, the peptides 542–566, 547–571, 660–677 and 597–614 have been found to bind specifically to calmodulin.Together with previously predicted and synthesized calmodulin binding peptides four calmodulin binding regions have been characterized on each the and subunits. It can be concluded that endogenous calmodulin can bind to two calmodulin binding regions in as well as to two regions in and . Exogenous calmodulin can bind to two regions in and in . A binding stoichiometry of 0.8mol of calmodulin/ protomer of phosphorylase kinase has been determined by inhibiting the ubiquitination of calmodulin with phosphorylase kinase. Phosphorylase kinase is half maximally activated by 23nM calmodulin which is in the affinity range of calmodulin binding peptides from to calmodulin. Therefore, binding of exogenous calmodulin to activates the enzyme. A model for switching endogenous calmodulin between , and and modulation of ATP binding to as well as Mg2+/ADP binding to by calmodulin is presented. 相似文献
57.
Comparison of the crystal structures of genetically engineered human manganese superoxide dismutase and manganese superoxide dismutase from Thermus thermophilus: differences in dimer-dimer interaction. 下载免费PDF全文
U. G. Wagner K. A. Pattridge M. L. Ludwig W. C. Stallings M. M. Werber C. Oefner F. Frolow J. L. Sussman 《Protein science : a publication of the Protein Society》1993,2(5):814-825
The three-dimensional X-ray structure of a recombinant human mitochondrial manganese superoxide dismutase (MnSOD) (chain length 198 residues) was determined by the method of molecular replacement using the related structure of MnSOD from Thermus thermophilus as a search model. This tetrameric human MnSOD crystallizes in space group P2(1)2(1)2 with a dimer in the asymmetric unit (Wagner, U.G., Werber, M.M., Beck, Y., Hartman, J.R., Frolow, F., & Sussman, J.L., 1989, J. Mol. Biol. 206, 787-788). Refinement of the protein structure (3,148 atoms with Mn and no solvents), with restraints maintaining noncrystallographic symmetry, converged at an R-factor of 0.207 using all data from 8.0 to 3.2 A resolution and group thermal parameters. The monomer-monomer interactions typical of bacterial Fe- and Mn-containing SODs are retained in the human enzyme, but the dimer-dimer interactions that form the tetramer are very different from those found in the structure of MnSOD from T. thermophilus. In human MnSOD one of the dimers is rotated by 84 degrees relative to its equivalent in the thermophile enzyme. As a result the monomers are arranged in an approximately tetrahedral array, the dimer-dimer packing is more intimate than observed in the bacterial MnSOD from T. thermophilus, and the dimers interdigitate. The metal-ligand interactions, determined by refinement and verified by computation of omit maps, are identical to those observed in T. thermophilus MnSOD. 相似文献
58.
59.
60.
Bettina Schmidt Thomas Tradler Jens-U. Rahfeld Birgit Ludwig Bunty Jain Karlheinz Mann K. Peter Rücknagel Bernhard Janowski Angelika Schierhorn Gerhard Küllertz Jörg Hacker Gunter Fischer 《Molecular microbiology》1996,21(6):1147-1160
Legionella pneumophila is the causative agent of a severe form of pneumonia in humans (Legionnaires’disease). A major virulence factor, the Mip protein (FK506-binding protein, FKBP25mem), belongs to the enzyme family of peptidyl-prolyl cis/trans isomerases (PPIases). Here we show that L. pneumophila Philadelphia I possesses an additional cytoplasmic PPiase at a level of enzyme activity comparable to that of FKBP25mem. The N-terminal amino acid sequence of the purified protein was obtained by Edman degradation and showed that the protein is a member of the cyclophilin family of PPIases. The Icy gene (Legionella cycophn) was cloned and sequenced. It encodes a putative 164-amino-acid protein with a molecular mass of 17 968 Da called L. pneumophila cyclophilin 18 (L. p. Cyp18). Amino acid sequence comparison displays considerable similarity to the cytoplasmic and the periplasmic cyclophilins of Escherichia coll with 60.5% and 51.5% identity, respectively. The substrate specificity and inhibition by cyclosporin A revealed a pattern that is typically found for other bacterial cyclophilins. An L. pneumophila Cyp18 derivative with a 19-amino-acid polypeptide extension including a 6-histi-dine tag and an enterokinase cleavage site exhibits 相似文献