全文获取类型
收费全文 | 3909篇 |
免费 | 280篇 |
国内免费 | 302篇 |
出版年
2024年 | 4篇 |
2023年 | 34篇 |
2022年 | 65篇 |
2021年 | 101篇 |
2020年 | 103篇 |
2019年 | 118篇 |
2018年 | 130篇 |
2017年 | 119篇 |
2016年 | 117篇 |
2015年 | 115篇 |
2014年 | 199篇 |
2013年 | 359篇 |
2012年 | 166篇 |
2011年 | 200篇 |
2010年 | 175篇 |
2009年 | 188篇 |
2008年 | 213篇 |
2007年 | 204篇 |
2006年 | 191篇 |
2005年 | 199篇 |
2004年 | 154篇 |
2003年 | 151篇 |
2002年 | 136篇 |
2001年 | 105篇 |
2000年 | 85篇 |
1999年 | 90篇 |
1998年 | 75篇 |
1997年 | 94篇 |
1996年 | 74篇 |
1995年 | 64篇 |
1994年 | 44篇 |
1993年 | 46篇 |
1992年 | 39篇 |
1991年 | 31篇 |
1990年 | 24篇 |
1989年 | 29篇 |
1988年 | 19篇 |
1987年 | 20篇 |
1986年 | 22篇 |
1985年 | 42篇 |
1984年 | 33篇 |
1983年 | 21篇 |
1982年 | 26篇 |
1981年 | 17篇 |
1980年 | 13篇 |
1979年 | 16篇 |
1978年 | 5篇 |
1977年 | 3篇 |
1976年 | 5篇 |
1974年 | 6篇 |
排序方式: 共有4491条查询结果,搜索用时 281 毫秒
81.
Environmental risk assessment of releases of transgenic plants containing virus-derived inserts 总被引:2,自引:0,他引:2
David J. Robinson 《Transgenic research》1996,5(5):359-362
Sequences derived from the genomes of plant viruses are being used to provide virus resistance in transgenic crop plants. Although the environmental hazards associated with the release of such plants have been discussed widely, it has not been possible to reach generally acceptable conclusions about their safety. A case-by-case approach to the risk assessment of real examples is recommended as a means of building up confidence and of indicating areas of uncertainty. A logical framework for risk assessment is suggested, a key feature of which is identification of the viruses in the release environment that may infect the transgenic plants. Each of these is considered in relation to each of the three main classes of hazard (transcapsidation, recombination and synergism), and the risk associated with each event is analysed. 相似文献
82.
Yoichi Honda Toshikazu Irie Mina Atsuji Takashi Watanabe Masaaki Kuwahara 《Mycoscience》1996,37(4):459-461
To develop a dominant genetic marker inPleurotus ostreatus, mutant strains resistant to a carboxin-derived fungicide, flutolanil, were isolated. These mutants included strains which
showed resistance to 50-fold higher concentration of fluotolanil than the wild-type strain, even after successive cultivations
in the absence of the drug. Dominance of the phenotype was confirmed by back-crossing between the resistant and wild-type
monokaryons. The flutolanilresistance was also shown to be stably inherited by the basidiospore-derived progenies of the mutant
strains. 相似文献
83.
During the past decade proteases have been widely used as catalysts in peptide synthesis. Unfortunately, they are not ideal ligases. Enzymatic peptide synthesis in frozen aqueous systems has been developed as an approach towards the suppression of competitive reactions. This paper summarizes reports concerning the behaviour of non-enzymatic as well as of enzyme-catalysed reactions when the reaction mixture is frozen. The advantages of freezing the reaction mixture in serine and cysteine protease-catalysed peptide synthesis, the influence of modified reaction conditions and the possible reasons for the yield-increasing effect of freezing are discussed. 相似文献
84.
Elín Gudmundsdóttir Rémi Spilliaert Qing Yang Charles S. Craik Jón B. Bjarnason Agusta Gudmundsdóttir 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1996,113(4):795-801
The cDNAs encoding two different Atlantic cod elastases have been isolated and sequenced. The predicted amino acid sequences revealed two preproelastases, consisting of a signal peptide, an activation peptide and a mature enzyme of 242 and 239 amino acids. Amino acid sequence identity between the two cod elastases was 60.1% and identity with mammalian elastases ranged from 50–64%. The two cod elastases contain all the major structural features common to serine proteases, such as the catalytic triad His57, Asp102 and Ser195. Both cod elastases have a high content of methionine, consistent with previous findings in psychrophilic fish enzymes. 相似文献
85.
Chang-Jen Huang †Chien-Chang Chen Hsiu-Jane Chen †Fore-Lien Huang ‡ Geen-Dong Chang 《Journal of neurochemistry》1995,64(4):1715-1720
Abstract: Two isoforms of a protease inhibitor of the serpin family (p62) have been purified from bighead carp perimeningeal fluid. Both isoforms migrate with an apparent molecular mass of 62 kDa on reducing and nonreducing sodium dodecyl sulfate-polyacrylamide gels. Both proteins inhibited the activities of bovine trypsin, bovine chymotrypsin, and porcine pancreatic elastase. They also formed complexes with these proteases that were resistant to sodium dodecyl sulfate treatment. p62 exists in the extracts of all tissues examined, including brain, head kidney, kidney, liver, muscle, ovary, pituitary, and spleen. It is also present in serum, ovarian fluid, and milt as well as perimeningeal fluid. The protease inhibitor is a glycoprotein, and its carbohydrate moiety could be removed by endoglycosidase F. Because p62 resembles mammalian α1 -antitrypsin in many aspects, it is likely a fish equivalent of α1 -antitrypsin. 相似文献
86.
Changes in the polypeptide composition of chloroplasts were investigated during germination of green spores of the fern Osmunda japonica . The polypeptide composition of chloroplasts was appreciably changed during a germination time course of 48 h. Levels of five polypeptides with apparent molecular masses of 47, 44, 42, 22 and 18.5 kDa in the soluble fraction of chloroplasts and three polypeptides with molecular masses of 24, 22 and 15 kDa in the thylakoid membranes decreased during germination. In contrast, no decrease of chloroplast polypeptides was observed in the spores incubated with cycloheximide for 48 h. A new 22-kDa protein was isolated from thylakoid membranes of spores and the amino-terminal sequence of the purified protein was determined. High levels of alanine and glycine were found in the basic protein (pl > 10.3). This protein, with a native molecular mass of 80 kDa, was characterized by a subunit band observed at a molecular mass of 22 kDa on SDS-PAGE and by the disappearance of the band during spore germination. Protease activity against the 22-kDa protein was observed in an extract prepared from chloroplasts of quiescent spores. A hypothetical cytosolic proteinaceous factor is implicated in the regulation of protein degradation in chloroplasts. 相似文献
87.
Genetic control of PI and GC variants in the American Mink 总被引:1,自引:0,他引:1
Genetic polymorphism of the serum α-protease inhibitor (PI) and group-specific component (GC) in minks was revealed using one-dimensional polyacrylamide gel electrophoresis and immunoblotting. Two codominant alleles were identified at each of the two loci. The data ruled out the possibility of any linkage between the PI, GC and the coat colour gene Crystal ( Cr ). 相似文献
88.
Abraham J. Koster Jochen Walz Andrei Lupas Wolfgang Baumeister 《Molecular biology reports》1995,21(1):11-20
The 26S proteasome is the central protease of the ubiquitin-dependent pathway of protein degradation. The molecule has a molecular mass of approximately 2000 kD and has a highly conserved structure in eukaryotes. The 26S proteasome is formed by a barrel-shaped 20S core complex and two polar 19S complexes. The 20S complex has C2 symmetry and is formed by four seven-membered rings of which the outer rings (-type subunits) are rotated by 25.7° relative to the inner rings while the inner rings (-type subunits) are in register. From a comparison of the activity and regulation of the 26S and 20S particles it can be deduced that the 20S particle contains the protease activity while the 19S complex contains isopeptidase, ATPase and protein unfolding activities. In this article we describe the structures of various proteasome complexes as determined by electron microscopy and discuss structural implications of their subunit sequences. 相似文献
89.
The authors established apoptosis resistant COS–1, myeloma, hybridoma, and Friend leukemia cell lines by genetically engineering cells, aiming at more efficient protein production by cell culture. COS–1 cells, which are most widely used for eukariotic gene expression, were transfected with human bcl–2 gene. Both bcl–2 and mock transfected COS–1 cells were cultured at low (0.2%) serum concentration for 9 days. The final viable cell number of the bcl–2 transfected cells was ninefold of that of the mock transfectants. Both bcl–2 and mock transfectants were further transfected with the vector pcDNA- containing SV40 ori and immunoglobulin gene for transiently expressing protein. The bcl–2 expressing COS–1 cells produced more protein than the mock transfected COS–1 cells after 4 days posttransfection.Mouse myeloma p3-X63-Ag.8.653 cells, which are widely used as the partner for preparing hybridoma, and hybridoma 2E3 cells were transfected with human bcl–2 gene. Both bcl–2 transfected myeloma and hybridoma survived longer than the corresponding original cells in batch culture. The bcl–2 transfected 2E3 cells survived 2 to 4 four days longer in culture, producing 1.5- to 4-fold amount of antibody in comparison with the mock transfectants.Coexpression of bag–1 with bcl–2 improved survival of hybridoma 2E3 cells more than bcl–2 expression alone. The bag–1 and bcl–2 coexpressing cells produced more IgG than the the cells expressing bcl–2 alone.Apoptosis of Friend murine erythroleukemia(F-MEL) cells was suppressed with antisense c-jun expression. The antisense c-jun expressing cells survived 16 days at non-growth state. 相似文献
90.
During the production by mammalian cells of recombinant factor VIII from which the B domain was deleted (rFVIII), proteolytic
cleavages in the C-terminal part of the heavy chain were observed (Kjalke et al., 1995). By radioactive pulse labelling it
was investigated whether the cleavages took place inside the cells during protein synthesis or after release in the medium.
The rFVIII-producing CHO (Chinese hamster ovary) cells were cultured in the presence of 35S-methionine and then the cell lysate and the conditioned media were immunoprecipitated and analyzed by electrophoresis. By
pulse labelling and chasing for various time periods, it was shown that the cleavages only took place after secretion of the
protein from the cells. Adding cell lysate to uncleaved rFVIII caused cleavage of the heavy chain, as seen by loss of binding
to a monoclonal antibody specific for intact rFVIII, indicating that the cleavage was performed by proteinase(s) released
from the lysed cells. By incubating intact rFVIII with the multicatalytic proteinase (proteasome) present in cytoplasm and
nucleus of eukaryotic cells, loss of binding to the monoclonal antibody was observed. This indicates that the multicatalytic
proteinase, released from lysed rFVIII producing cells, could be responsible for the cleavage of rFVIII. Among several protease
inhibitors tested, only bacitracin was found to diminish the extent of cleavage. Phosphatidylserine also protected rFVIII
against cleavage, probably by binding to rFVIII.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献