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81.
Albert S. Reger Matthew P. Yang Shizuyo Koide-Yoshida Elaine Guo Shrenik Mehta Keizo Yuasa Alan Liu Darren E. Casteel Choel Kim 《The Journal of biological chemistry》2014,289(37):25393-25403
cGMP-dependent protein kinase (PKG)-interacting proteins (GKIPs) mediate cellular targeting of PKG isoforms by interacting with their leucine zipper (LZ) domains. These interactions prevent aberrant signaling cross-talk between different PKG isotypes. To gain detailed insight into isotype-specific GKIP recognition by PKG, we analyzed the type II PKG leucine zipper domain and found that residues 40–83 dimerized and specifically interacted with Rab11b. Next, we determined a crystal structure of the PKG II LZ-Rab11b complex. The PKG II LZ domain presents a mostly nonpolar surface onto which Rab11b docks, through van der Waals interactions. Contact surfaces in Rab11b are found in switch I and II, interswitch, and the β1/N-terminal regions. This binding surface dramatically differs from that seen in the Rab11 family of interacting protein complex structures. Structural comparison with PKG Iα and Iβ LZs combined with mutagenic analysis reveals that GKIP recognition is mediated through surface charge interactions. 相似文献
82.
Kan Fujino Masayuki Horie Tomoyuki Honda Shoko Nakamura Yusuke Matsumoto Ivo M. B. Francischetti Keizo Tomonaga 《PloS one》2012,7(12)
Bornavirus, a non-segmented, negative-strand RNA viruses, is currently classified into several genetically distinct genotypes, such as Borna disease virus (BDV) and avian bornaviruses (ABVs). Recent studies revealed that bornavirus genotypes show unique sequence variability in the putative 5′ untranslated region (5′ UTR) of X/P mRNA, a bicistronic mRNA for the X protein and phosphoprotein (P). In this study, to understand the evolutionary relationship among the bornavirus genotypes, we investigated the functional interaction between the X and P proteins of four bornavirus genotypes, BDV, ABV genotype 4 and 5 and reptile bornavirus (RBV), the putative 5′ UTRs of which exhibit variation in the length. Immunofluorescence and immunoprecipitation analyses using mammalian and avian cell lines revealed that the X proteins of bornaviruses conserve the ability to facilitate the export of P from the nucleus to the cytoplasm via interaction with P. Furthermore, we showed that inter-genotypic interactions may occur between X and P among the genotypes, except for X of RBV. In addition, a BDV minireplicon assay demonstrated that the X and P proteins of ABVs, but not RBV, can affect the polymerase activity of BDV. This study demonstrates that bornaviruses may have conserved the fundamental function of a regulatory protein during their evolution, whereas RBV has evolved distinctly from the other bornavirus genotypes. 相似文献
83.
Koji Ashizawa Etsuko Tamiya Keizo Okauchi Hisayoshi Nishiyama 《Animal reproduction science》1985,9(2):181-188
Fowl spermatozoa were incubated at 41°C in a supernatant removed from a 4-day culture medium of embryonic chick skeletal muscle cells. Their motility, as assessed at room temperature (20–25°C), was maintained for 48 h. Fertilizing ability was also retained for at least 24 h. In contrast, spermatozoa incubated in the fresh culture medium lost their motility and fertilizing ability rapidly. A filtrate of the 4-day culture medium, obtained by passing the fluid through an Amicon PM-10 ultrafiltration membrane, prolonged the motility of spermatozoa. These results suggested that a low molecular weight factor(s) (mol. wt. < 10 000) supplied by the cultured cells effectively prolonged the motility of spermatozoa. 相似文献
84.
Resistance of staphylococci to desiccation, dry heat, wet heat and ultraviolet irradiation was studied using 128 strains obtained from patients in our surgical clinics. Staphylococci of phage group I including type 81, particularly of type 80/81, showed significantly high death rates after desiccation, dry heating, and wet heating, and significantly low death rates after ultraviolet irradiation. In contrast, group-II strains responded inversely to these physical agents. The mean death rate of group-III strains approximated that of group I after desiccation, dry heating, and ultraviolet irradiation, but after wet heating it was lower than that of group II. The results indicate that transmissibility of staphylococci might be influenced to a great extent by physical factors in the environment outside the body. 相似文献
85.
Determination of
Ichiro Ono Keizo Matsuda Sachiko Kanno 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1997,692(2)
A simple, rapid and sensitive two column-switching high-performance liquid chromatographic (HPLC) method with ultraviolet detection at 210 nm has been developed for the determination of
(AY4166, I) and its seven metabolites in human plasma and urine. Measurements of I and its metabolites were carried out by two column-switching HPLC, because metabolites were classified into two groups according to their retention times. After purification of plasma samples using solid-phase extraction and direct dilution of urinary samples, I and each metabolite were injected into HPLC. The calibration graphs for plasma and urinary samples were linear in the ranges 0.1 to 10 μg ml−1 and 0.5 to 50 μg ml−1, respectively. Recoveries of I and its seven metabolites were over 88% by the standard addition method and the relative standard deviations of I and its metabolites were 1–6%. 相似文献
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86.
87.
The mechanism of the lytic action of prymnesin, a toxin produced by the alga, Prymnesium parvum, was studied using liposomes as a model membrane system. Prymnesin showed severe damage to liposomes containing cholesterol but did not affect those without cholesterol. The requirement of cholesterol for the susceptibility to prymnesin was much more strict than the reported requirement for the susceptibility to polyene antibiotics. The net charge on the membranes was shown not to be important for the reaction. 相似文献
88.
Sequence determination of a new parrot bornavirus‐5 strain in Japan: implications of clade‐specific sequence diversity in the regions interacting with host factors
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Ryo Komorizono Akiko Makino Masayuki Horie Tomoyuki Honda Keizo Tomonaga 《Microbiology and immunology》2016,60(6):437-441
In this study, the genome sequence of a new parrot bornavirus‐5 (PaBV‐5) detected in Eclectus roratus was determined. Phylogenetic analysis showed that the genus Bornavirus is divided into three major clades and that PaBV‐5 belongs to clade 2, which contains avian viruses that exhibit infectivity to mammalian cells. Sequence comparisons of the regions known to interact with host factors indicated that the clade 2 avian viruses possess sequences intermediate between the clade 1 mammalian viruses and the clade 3 avian viruses, suggesting that the identified regions might contribute to the differences in virological properties between the three clades. 相似文献
89.
James C. Campbell Jeong Joo Kim Kevin Y. Li Gilbert Y. Huang Albert S. Reger Shinya Matsuda Banumathi Sankaran Todd M. Link Keizo Yuasa John E. Ladbury Darren E. Casteel Choel Kim 《The Journal of biological chemistry》2016,291(11):5623-5633
Membrane-bound cGMP-dependent protein kinase (PKG) II is a key regulator of bone growth, renin secretion, and memory formation. Despite its crucial physiological roles, little is known about its cyclic nucleotide selectivity mechanism due to a lack of structural information. Here, we find that the C-terminal cyclic nucleotide binding (CNB-B) domain of PKG II binds cGMP with higher affinity and selectivity when compared with its N-terminal CNB (CNB-A) domain. To understand the structural basis of cGMP selectivity, we solved co-crystal structures of the CNB domains with cyclic nucleotides. Our structures combined with mutagenesis demonstrate that the guanine-specific contacts at Asp-412 and Arg-415 of the αC-helix of CNB-B are crucial for cGMP selectivity and activation of PKG II. Structural comparison with the cGMP selective CNB domains of human PKG I and Plasmodium falciparum PKG (PfPKG) shows different contacts with the guanine moiety, revealing a unique cGMP selectivity mechanism for PKG II. 相似文献
90.
Yamashita A Nakanishi H Suzuki H Kamata R Tanaka K Waku K Sugiura T 《Biochimica et biophysica acta》2007,1771(9):1202-1215
1-acyl-sn-glycero-3-phosphate (AGP) acyltransferases (AGPAT) are involved in de novo biosynthesis of glycerolipids, such as phospholipids and triacylglycerol. Alignment of amino acid sequences from AGPAT, sn-glycerol-3-phosphate acyltransferase, and dihydroxyacetonephosphate acyltransferase reveals four regions with strong homology (acyltransferase motifs I-IV). The invariant amino acids within these regions may be part of a catalytically important site in this group of acyl-CoA acyltransferases. However, in human AGPAT1 a transmembrane domain is predicted to separate motif I on the cytosolic side from motifs II-III on the lumenal side, with motif IV near surface of the membrane. The topology of motifs I and III was confirmed by experiments with recombinant AGPAT1 containing potential glycosylation site near the motifs. This topology conflicts with the expectation that catalytically important sites are near one another, raising questions of whether the acyltransferase motifs really are important for AGPAT catalysis, and how substrates access motifs II-III on the lumenal side of the endoplasmic reticulum membrane. Using human AGPAT1 as a model, we have examined the catalytic roles of highly conserved residues in the four acyltransferase motifs by site-directed mutagenesis. Modifications of the sidechain structures of His104, Asp109, Phe146, Arg149, Glu178, Gly179, Thr180, Arg181 and Ile208 all affected AGPAT1 activity, indicating that the acyltransferase motifs indeed are important for AGPAT catalysis. In addition, we examined substrate accessibility to the catalytic domain of human AGPAT1 using a competition assay. Lysophosphatidic acid (LPA) with fatty acid chains shorter than 10 carbons did not access the catalytic domain, suggesting that LPA hydrophobicity is important. In contrast, short chain acyl-CoAs did access the catalytic domain but did not serve as the second substrate. These results suggest that motifs II and III are involved in LPA binding and motifs I and IV are involved in acyl-CoA binding. 相似文献