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251.
Hideaki Yukawa Yasurou Kurusu Mitsunobu Shimazu Hisashi Yamagata Masato Terasawa 《Journal of industrial microbiology & biotechnology》1988,2(6):323-328
Summary In anEscherichia coli K-12 strain (trpA trpE tnaA) cultured in LB broth without selective pressure, a pBR322 derivative containing the gene for tryptophan synthase (pBR322-trpBA) was found to be unstable. After 70 cell-number doublings, only 50% of the host cells retained the gene for ampicillin resistance (Apr). Insertion of the mini-F fragment of F factor DNA into this plasmid could effectively reduce the plasmid loss. Partial derepression of the tryptophan promotor-operator by 3-indopleacrylic acid further decreased the stability of the pBR322-trpBA but not that of the mini-F inserted plasmid (pBR322F-trpBA) The vector pBR322F-trpBA could be maintained at high copy number in the culture after 100 generations of growth; the culture was able to overproduce tryptophan synthase in the presence of 3-indoleacrylic acid.l-Tryptophan was produced from indole andl-serine using andE. coli host transformed with.pBR322F-trpBA DNA. After 8 h of incubation, the expression level was approximately 180 g/l. 相似文献
252.
The cadherin superfamily is a large protein family with diverse structures and functions. Because of this diversity and the growing biological interest in cell adhesion and signaling processes, in which many members of the cadherin superfamily play a crucial role, it is becoming increasingly important to develop tools to manage, distribute and analyze sequences in this protein family. Current profile and motif databases classify protein sequences into a broad spectrum of protein superfamilies, however to provide a more specific functional annotation, the next step should include classification of subfamilies of these protein superfamilies. Here, we present a tool that classified greater than 90% of the proteins belonging to the cadherin superfamily found in the SWISS PROT database. Therefore, for most members of the cadherin superfamily, this tool can assist in adding more specific functional annotations than can be achieved with current profile and motif databases. Finally, the classification tool and the results of our analysis were integrated into a web-accessible database (http://calcium.uhnres. utoronto.ca/cadherin). 相似文献
253.
Calmodulin-regulated serine/threonine kinases (CaM kinases) play crucial roles in Ca2+-dependent signaling transduction pathways in eukaryotes. Despite having a similar overall molecular architecture of catalytic and regulatory domains, CaM kinases employ different binding modes for Ca2+/CaM recruitment which is required for their activation. Here we present a residual dipolar coupling (RDC)-based NMR approach to characterizing the molecular recognition of CaM with five different CaM kinases. Our analyses indicate that CaM kinase I and likely IV use the same CaM binding mode as myosin light chain kinase (1-14 motif), distinct from those of CaM kinase II (1-10 motif) and CaM kinase kinase (1-16- motif). This NMR approach provides an efficient experimental guide for homology modeling and structural characterization of CaM-target complexes. 相似文献
254.
Improved three-dimensional1H−13C−1H correlation spectroscopy of a13C-labeled protein using constant-time evolution 总被引:1,自引:0,他引:1
Summary An improved version of the three-dimensional HCCH-COSY NMR experiment is described that correlates the resonances of geminal and vicinal proton pairs with the chemical shift of the13C nucleus attached to one of the protons. The experiment uses constant-time evolution of transverse13C magnetization which optimizes transfer of magnetization and thus improves the sensitivity of the experiment over the original scheme. The experiment is demonstrated for calmodulin complexed with a 26-residue peptide comprising the binding site of skeletal muscle myosin light chain kinase. 相似文献
255.
We investigated the existence and possible role of cytosolic phospholipase A2 (cPLA2) in rat decidualized uteri. PLA2 activity in the cytosol of a decidualized uterine horn, induced by intraluminal oil infusion, was significantly higher than that in contralateral intact horn. The activity was almost completely depressed by cPLA2 inhibitors including arachidonyl trifluoromethyl ketone (ATK). The immunoreactive signals for cPLA2 were intense in decidua and glandular epithelial cells. In vivo administration of ATK (0.1-100 microg) caused a dose-dependent inhibition of decidualization. These results show the presence of cPLA2 and its probable implication in decidualization in rat uterus. 相似文献
256.
We investigated the mechanism by which rat luteal cells produce prostaglandin F(2 alpha) (PGF(2 alpha)) and its relevance to cell death in vitro. Treatment with progesterone (P4) of dispersed luteal cells prepared from rats on day 9 of pseudopregnancy caused dose-dependent inhibition of PGF(2 alpha) secretion. Cytokines, tumor necrosis factor alpha (TNFalpha) or interferon gamma (IFN gamma) alone had no or modest regulatory effects. Arachidonyl trifluoromethyl ketone (AACOCF(3)), a specific group IVA phospholipase A(2) inhibitor, depressed both basal and cytokine-regulated PGF(2 alpha) production. A combination of TNFalpha and IFN gamma stimulated PGF(2 alpha) synthesis and cytotoxicity (both, P<0.05). Agonistic anti-Fas antibody challenge caused a significant cytotoxic effect but without affecting PGF(2 alpha) production. The present data suggest that P4 inhibits and TNFalpha and IFN gamma cooperatively stimulate PGF(2 alpha) release by rat luteal cells. They also suggest that luteal cell death induced by TNFalpha/IFN gamma and Fas stimulation seems to occur via distinct signaling pathways involving PGF(2 alpha) production. 相似文献
257.
Alain A. Vertès Yoko Asai Masayuki Inui Miki Kobayashi Yasurou Kurusu Hideaki Yukawa 《Molecular & general genetics : MGG》1994,245(4):397-405
The Corynebacterium glutamicum insertion sequence IS31831 was used to construct two artificial transposons: Tn31831 and miniTn31831. The transposition vectors were based on a gram-negative replication origin and do not replicate in coryneform bacteria. Strain Brevibacterium flavum MJ233C was mutagenized by miniTn31831 at an efficiency of 4.3 x 104 mutants per microgram DNA. Transposon insertions occurred at different locations on the chromosome and produced a variety of mutants. Auxotrophs could be recovered at a frequency of approximately 0.2%. Transposition of IS31831 derivatives led not only to simple insertion, but also to cointegrate formation (5%). No multiple insertions were observed. Chromosomal loci of B. flavum corresponding to auxotrophic and pigmentation mutants could be rescued in Escherichia coli, demonstrating that these transposable elements are useful genetic tools for studying the biology of coryneform bacteria. 相似文献
258.
Akaboshi Mitsuhiko Tanaka Yoshiko Nakano Yukihiro Kawai Kenichi Fujii Noriko 《Origins of life and evolution of the biosphere》2000,30(1):25-32
Catalytic action of rare earth element, Ce(IV) to hydrolyzephosphomonoester bonds was confirmed. This effect wasconsidered to suppress abiotic synthesis ofnucleotides and nucleic acids in the primitive sea,and hence the origin of life. However, we found thatthe presence of proteins, especially albumin, stronglyinhibited the catalytic action of Ce(IV). Thisfinding was supported by preferential binding of rare earthelements (REEs) to proteins which was revealed using the radioisotopes of these REEs. Consequently, if a large amount ofproteins was synthesized in the primitive sea, abioticsynthesis of phosphomonoester compounds, and hencenucleic acids, might have been possible. 相似文献
259.
Toyoto Hijiya Keizo Yamashita Mitsuhiko Kojima Yumiko Uchida Satoshi Katayama Takayoshi Torii 《Nucleosides, nucleotides & nucleic acids》2013,32(4-5):653-654
Abstract An economical synthesis of famciclovir from N-2-acetyl-7-benzylguanine by a novel regioselective alkylation with the diester cyclopropane compound was developed. 相似文献
260.