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排序方式: 共有121条查询结果,搜索用时 281 毫秒
1.
Bernardo Celda Clelia Biamonti Maria Jose Arnau Roberto Tejero Gaetano T. Montelione 《Journal of biomolecular NMR》1995,5(2):161-172
Summary A large portion of the 13C resonance assignments for murine epidermal growth factor (mEGF) at pH 3.1 and 28°C has been determined at natural isotope abundance. Sequence-specific 13C assignments are reported for 100% of the assignable C, 96% of the C, 86% of the aromatic and 70% of the remaining peripheral aliphatic resonances of mEGF. A good correlation was observed between experimental and back-calculated C chemical shifts for regions of regular -sheet structure. These assignments also provide the basis for interpreting 1H–13C heteronuclear NOE (HNOE) values in mEGF at natural isotope abundance. Some of the backbone polypeptide segments with high internal mobility, indicated by these 1H–13C HNOE measurements, correlate with locations of residues involved in the putative mEGF-receptor binding site. Using four families of mEGF structures obtained over the last few years, we demonstrate that standard deviations between experimental and back-calculated C values can be used to monitor the refinement of this protein's structure, particularly for -sheet regions. Improved agreement between calculated and observed values of C is correlated with other measures of structure quality, including lowered values of residual constraint violations and more negative values of conformational energy. These results support the view that experimental conformation-dependent chemical shifts, C, can provide a reliable source of information for monitoring the process of protein structure refinement and are potentially useful restraints for driving the refinement.Abbreviations HSQC
heteronuclear single-quantum coherence spectroscopy
- PFG
pulsed-field gradient
- TOCSY
1H-1H total correlation spectroscopy
- EGF
epidermal growth factor
- mEGF
murine EGF
- hEGF
human EGF
- hTGF
human type- transforming growth factor
- DIPSI
spm-locking pulse sequence
- NOE
nuclear Overhauser effect
- HNOE
heteronuclear Overhauser effect 相似文献
2.
Anke Burmester Anke Wöstemeyer Jose Arnau Johannes Wöstemeyer 《Molecular & general genetics : MGG》1992,235(2-3):166-172
Summary A series of new vectors for the model zygomycete Absidia glauca was constructed on the basis of the structural neomycin resistance (Neor) gene controlled by the promoter of the gene for elongation factor 1 (TEF). In order to select for transformed colonies with a stable Neor phenotype, spores from primary transformants were pooled and grown for two sporulation cycles under non-selective conditions. Southern blot analysis of DNA from single spore isolates originating from independent transformant pools allowed the identification of two autonomously replicating plasmids. Retransformation of Escherichia coli and restriction analysis of the two plasmids provided evidence for spontaneous in vivo insertion of a new DNA element (SEG1) from the A. glauca genome. The inserted regions in both plasmids are essentially identical and do not represent repetitive DNA. Compared with other autonomously replicating vectors, these SEG1-containing plasmids are mitotically extremely stable and are passed on to the vegetative spore progeny of a retransformed A. glauca strain. We assume that SEG1 contains structural elements involved in partitioning and stable segregation of plasmids. For the construction of stable transformants of A. glauca, the SEG1 element may be regarded as a major breakthrough, because stabilization of transformed genetic traits by integration is difficult to achieve in all mucoraceous fungi and all known replicating plasmids are mitotically unstable. 相似文献
3.
Molecular Dynamics Simulations of Rhodopsin Point Mutants at the Cytoplasmic Side of Helices 3 and 6
Arnau Cordomí Eva Ramon Pere Garriga Juan J. Perez 《Journal of biomolecular structure & dynamics》2013,31(6):573-587
Abstract The present work reports on a structural analysis carried out through different computer simulations of a set of rhodopsin mutants with differential functional features in regard to the wild type. Most of these mutants, whose experimental features had previously been reported [Ramon et al. J Biol Chem 282, 14272–14282 (2007)], were designed to perturb a network of electrostatic interactions located at the cytoplasmic sides of transmembrane helices 3 and 6. Geometric and energetic features derived from the detailed analysis of a series of molecular dynamics simulations of the different rhodopsin mutants, involving positions 134(3.49), 247(6.30), and 251(6.34), suggest that the protein structure is sensitive to these mutations through the local changes induced that extend further to the secondary structure of neighboring helices and, ultimately, to the packing of the helical bundle. Overall, the results obtained highlight the complexity of the analyzed network of electrostatic interactions where the effect of each mutation on protein structure can produce rather specific features. 相似文献
4.
Coronado Tomás M. Mir Arnau Rosselló Francesc Valiente Gabriel 《Journal of mathematical biology》2019,79(3):1105-1148
Journal of Mathematical Biology - We define a new balance index for rooted phylogenetic trees based on the symmetry of the evolutive history of every set of 4 leaves. This index makes sense for... 相似文献
5.
José Arnau Antonio Ortiz Juan C. Gomez-Fernández Francisco J. Murillo Santiago Torres-Martínez 《FEMS microbiology letters》1988,51(1):37-40
Abstract We describe here fusion between phospholipid vesicles (liposomes) and protoplasts to the fungus Phycomyces blakesleeanus . Both 6-carboxyfluorescein and the kanamycin resistance harboured by the plasmid have been transferred from liposomes to protoplasts of Phycomyces by the fusion technique. 相似文献
6.
Mucor circinelloides is being investigated as a possible host for the production of heterologous proteins. Thus, the environmental conditions defining the physiology and morphology of this dimorphic fungus have been investigated in submerged batch cultivation. The optimal conditions for growth of each form have been defined. Pure cultures of the multi-polar budding yeast form could be obtained under anaerobic conditions (with 70% N2/30% CO2 or 100% N2 as the sparge gas and without aeration). The highest maximum specific growth rate (0.30 h(-1)) was obtained in anaerobic cultivation, the yield of biomass on glucose (Y(SX)) was 0.12 (c-mole basis). A high maximum specific growth rate was obtained when the organism grew as the filamentous form under aerobic conditions (0.25 h(-1)), with a Y(SX) of 0.24 (c-mole basis). The maximum specific growth rates achieved are comparable to most industrial filamentous fungi under similar growth conditions. High levels of ethanol were observed with all growth conditions. The overriding effector of morphological development was found to be oxygen. In batch cultures it was therefore possible to induce the dimorphic shift by controlling the influent gas atmosphere. A specific growth rate of 0.19 h(-1) was maintained during the shift from the yeast to the filamentous form. 相似文献
7.
Carmen S. M. Yong Janelle Sharkey Belinda Duscio Ben Venville Wei-Zen Wei Richard F. Jones Clare Y. Slaney Gisela Mir Arnau Anthony T. Papenfuss Jan Schr?der Phillip K. Darcy Michael H. Kershaw 《PloS one》2015,10(9)
The development of antigen-targeted therapeutics is dependent on the preferential expression of tumor-associated antigens (TAA) at targetable levels on the tumor. Tumor-associated antigens can be generated de novo or can arise from altered expression of normal basal proteins, such as the up-regulation of human epidermal growth factor receptor 2 (Her2/ErbB2). To properly assess the development of Her2 therapeutics in an immune tolerant model, we previously generated a transgenic mouse model in which expression of the human Her2 protein was present in both the brain and mammary tissue. This mouse model has facilitated the development of Her2 targeted therapies in a clinically relevant and suitable model. While heterozygous Her2+/- mice appear to develop in a similar manner to wild type mice (Her2-/-), it has proven difficult to generate homozygous Her2+/+ mice, potentially due to embryonic lethality. In this study, we performed whole genome sequencing to determine if the integration site of the Her2 transgene was responsible for this lethality. Indeed, we report that the Her2 transgene had integrated into the Pds5b (precocious dissociation of sisters) gene on chromosome 5, as a 162 copy concatemer. Furthermore, our findings demonstrate that Her2+/+ mice, similar to Pds5b-/- mice, are embryonic lethal and confirm the necessity for Pds5b in embryonic development. This study confirms the value of whole genome sequencing in determining the integration site of transgenes to gain insight into associated phenotypes. 相似文献
8.
Leire Aguinagalde Bruno Corsini Arnau Domenech Mirian Domenech Jordi Cámara Carmen Ardanuy Ernesto García Josefina Li?ares Asunción Fenoll Jose Yuste 《PloS one》2015,10(9)
Capsular switching allows pre-existing clones of Streptococcus pneumoniae expressing vaccine serotypes to escape the vaccine-induced immunity by acquisition of capsular genes from pneumococci of a non-vaccine serotype. Here, we have analysed the clonal composition of 492 clinical isolates of serotype 11A causing invasive disease in Spain (2000–2012), and their ability to evade the host immune response. Antibiograms, serotyping and molecular typing were performed. The restriction profiles of pbp2x, pbp1a and pbp2b genes were also analysed. Interaction with the complement components C1q, C3b, C4BP, and factor H was explored whereas opsonophagocytosis assays were performed using a human cell line differentiated to neutrophils. Biofilm formation and the polymorphisms of the major autolysin LytA were evaluated. The main genotypes of the 11A pneumococci were: ST62 (447 isolates, 90.6%), followed by ST6521 (35 isolates, 7.3%) and ST838 (10 isolates, 2.1%). Beta lactam resistant serotype 11A variants of genotypes ST838 and ST6521 closely related to the Spain9V-ST156 clone were first detected in 2005. A different pattern of evasion of complement immunity and phagocytosis was observed between genotypes. The emergence of one vaccine escape variant of Spain9V-ST156 (ST652111A), showing a high potential to avoid the host immune response, was observed. In addition, isolates of ST652111A showed higher ability to produce biofilms than ST83811A or ST6211A, which may have contributed to the emergence of this PEN-resistant ST652111A genotype in the last few years. The emergence of penicillin-resistant 11A invasive variants of the highly successful ST156 clonal complex merits close monitoring. 相似文献
9.
Miriam Masià-Balagué Ismael Izquierdo Georgina Garrido Arnau Cordomí Laura Pérez-Benito Nichol L. G. Miller David D. Schlaepfer Véronique Gigoux Anna M. Aragay 《The Journal of biological chemistry》2015,290(24):15197-15209
The guanine nucleotide exchange factor Rgnef (also known as ArhGEF28 or p190RhoGEF) promotes colon carcinoma cell motility and tumor progression via interaction with focal adhesion kinase (FAK). Mechanisms of Rgnef activation downstream of integrin or G protein-coupled receptors remain undefined. In the absence of a recognized G protein signaling homology domain in Rgnef, no proximal linkage to G proteins was known. Utilizing multiple methods, we have identified Rgnef as a new effector for Gα13 downstream of gastrin and the type 2 cholecystokinin receptor. In DLD-1 colon carcinoma cells depleted of Gα13, gastrin-induced FAK Tyr(P)-397 and paxillin Tyr(P)-31 phosphorylation were reduced. RhoA GTP binding and promoter activity were increased by Rgnef in combination with active Gα13. Rgnef co-immunoprecipitated with activated Gα13Q226L but not Gα12Q229L. The Rgnef C-terminal (CT, 1279–1582) region was sufficient for co-immunoprecipitation, and Rgnef-CT exogenous expression prevented Gα13-stimulated SRE activity. A domain at the C terminus of the protein close to the FAK binding domain is necessary to bind to Gα13. Point mutations of Rgnef-CT residues disrupt association with active Gα13 but not Gαq. These results show that Rgnef functions as an effector of Gα13 signaling and that this linkage may mediate FAK activation in DLD-1 colon carcinoma cells. 相似文献
10.