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951.
952.
CA Petta A Faúndes TR Dunson M Ramos M DeLucio D Faúndes L Bahamondes 《Canadian Metallurgical Quarterly》1998,70(5):817-820
1. Microsomal P450 and peroxisomal fatty acid oxidation activities were studied in liver of rats after long-term ethanol consumption. 2. Ethanol increased the microsomal lauric acid omega-hydroxylation and the aminopyrine N-demethylation catalyzed by cytochrome P450. 3. Ethanol increased peroxisomal beta-oxidation of palmitoyl CoA and catalase activity in liver. 4. Both microsomal and peroxisomal activities behaved in a coordinate way in the liver of rats with long-term ethanol consumption. 5. These results would support a role of microsomal omega-hydroxylation and peroxisomal beta-oxidation of fatty acids in an extramitochondrial pathway of lipid oxidation in the liver. 相似文献
953.
TR Jones SE Webber MD Varney MR Reddy KK Lewis V Kathardekar H Mazdiyasni J Deal D Nguyen KM Welsh S Webber A Johnston DA Matthews WW Smith CA Janson RJ Bacquet EF Howland CL Booth SM Herrmann RW Ward J White CA Bartlett CA Morse 《Canadian Metallurgical Quarterly》1997,40(5):677-683
Six new diphenyl sulfoxide and five new diphenyl sulfones were designed, synthesized, and tested for their inhibition of human and Escherichia coli thymidylate synthase (TS) and of the growth of cells in tissue culture. The best sulfoxide inhibitor of human TS was 3-chloro-N-((3,4-dihydro-2-methyl-4-oxo-6-quinazolinyl)methyl)-4- (phenylsulfinyl)-N-(prop-2-ynyl)-aniline (7c) that had a Ki of 27 nM. No sulfone improved on TS inhibition by the previously reported 4-(N-((3,4-dihydro-2-methyl-6-quinazolinyl)methyl)-N-prop-2- ynylamino)phenyl phenyl sulfone (Ki = 12 nM). Nevertheless, one sulfone, 4-((2-chlorophenyl)sulfonyl)-N-((3,4-dihydro-2-methyl-4-oxo-6- quinazolinyl)methyl)-N-(prop-2-ynyl)aniline, was selected, on the basis of its inhibition of both TS and cell growth, for antitumor testing; it gave a 61% increase in life span to mice bearing the thymidino kinase-deficient L5178Y (TK-) lymphoma. A crystal structure of N-((3,4-dihydro-2-methyl-4-oxo-6-quinazolinyl)methyl)-4-((2- methylphenyl)sulfinyl)-N-(prop-2-ynyl)aniline complexed with E. coli TS was solved and revealed selective binding of one sulfoxide enantiomer. AMBER calculations showed that the enantioselection was due to asymmetric electrostatic effects at the mouth of the active site. In contrast, a similar crystal structure of the sulfoxide 7c, along with AMBER calculations, indicated that both enantiomers bound, but with different affinities. The side chain of Phe176 shifted in order to structurally accommodate the chlorine of the more weakly bound enantiomer. 相似文献
954.
High-level expression of bovine beta-lactoglobulin in Pichia pastoris and characterization of its physical properties 总被引:3,自引:0,他引:3
Kim TR; Goto Y; Hirota N; Kuwata K; Denton H; Wu SY; Sawyer L; Batt CA 《Protein engineering, design & selection : PEDS》1997,10(11):1339-1345
Bovine beta-lactoglobulin (BLG) variant A has been expressed in the
methylotropic yeast Pichia pastoris by fusion of the cDNA to the sequence
coding for the alpha-mating factor prepro-leader peptide from Saccharomyces
cerevisiae. P. pastoris Mut+ transformants were obtained by single
cross-over integration of the BLG-containing vector into the AOX1 locus. In
a fed-batch fermenter, a cell density of approximately 300 mg/ml was
achieved by controlled glycerol feeding for a total of 24 h. After 72 h of
methanol induction, the secreted BLG reached levels of > 1 g/l. The
secreted protein could be purified to homogeneity by ion- exchange
chromatography. Amino-terminal sequencing of the secreted BLG revealed that
the Glu-Ala spacer repeats inserted between the mature protein and the
alpha-factor prepro-leader were still present. The purified protein was
characterized by a number of methods, including CD spectroscopy,
guanidine-HCl unfolding, crystallization and two- dimensional 1H-NMR
spectroscopy. By all of these measures, the physical characteristics of
recombinant BLG were indistinguishable from those of the native purified
bovine BLG, making it useful as a model for protein folding and other
biophysical studies.
相似文献
955.
Recent studies have provided evidence to implicate involvement of the core oligosaccharide region of Pseudomonas aeruginosa lipopolysaccharide (LPS) in adherence to host tissues. To better understand the role played by LPS in the virulence of this organism, the aim of the present study was to clone and characterize genes involved in core biosynthesis. The inner-core regions of P. aeruginosa and Salmonella enterica serovar Typhimurium are structurally very similar; both contain two main chain residues of heptose linked to lipid A-Kdo2 (Kdo is 3-deoxy-D-manno-octulosonic acid). By electrotransforming a P. aeruginosa PAO1 library into Salmonella waaC and waaF (formerly known as rfaC and rfaF, respectively) mutants, we were able to isolate the homologous heptosyltransferase I and II genes of P. aeruginosa. Two plasmids, pCOREc1 and pCOREc2, which restored smooth LPS production in the waaC mutant, were isolated. Similarly, plasmid pCOREf1 was able to complement the Salmonella waaF mutant. Sequence analysis of the DNA insert of pCOREc2 revealed one open reading frame (ORF) which could code for a protein of 39.8 kDa. The amino acid sequence of the deduced protein exhibited 53% identity with the sequence of the WaaC protein of S. enterica serovar Typhimurium. pCOREf1 contained one ORF capable of encoding a 38.4-kDa protein. The sequence of the predicted protein was 49% identical to the sequence of the Salmonella WaaF protein. Protein expression by the Maxicell system confirmed that a 40-kDa protein was encoded by pCOREc2 and a 38-kDa protein was encoded by pCOREf1. Pulsed-field gel electrophoresis was used to determine the map locations of the cloned waaC and waaF genes, which were found to lie between 0.9 and 6.6 min on the PAO1 chromosome. Using a gene-replacement strategy, we attempted to generate P. aeruginosa waaC and waaF null mutants. Despite multiple attempts to isolate true knockout mutants, all transconjugants were identified as merodiploids. 相似文献
956.
957.
958.
The structures of the cyclic hexapeptide cyclo(-Gly-Tyr-Val-Pro-Met-Leu-) (1) and its phosphotyrosyl (pTyr) derivative cyclo[-Gly-Tyr(PO3H2)-Val-Pro-Met-Leu-] (2), designed as constrained models of a sequence that interacts with the src homology 2 (SH2) region of the p85 subunit of phosphatidylinositol-3-OH kinase (PI-3 kinase), were studied in methanol/water solutions by 500 MHz nmr spectroscopy. Compound 1 was found to exist as a 2:1 mixture of isomers about the Val-Pro bond (trans and cis prolyl) between 292-330 K in 75% CD3O(D,H)/(D,H)2O solutions. A third species of undetermined structure (ca. 5%) was also observed. Compound 2, a model of phosphorylated peptide ligand that binds to the PI-3 kinase SH2 domain, exhibited similar conformational isomerism. When either compound was dissolved in pure solvent [i.e., 100% CD3O(H,D) or (H,D)2O] the ratio of cis to trans isomers was ca 1:1. A battery of one- and two-dimensional nmr experiments at different temperatures and solvent compositions allowed a complete assignment of both the cis and trans forms of 1 and indicated the trans compound to be the major isomer. The spectral properties of the phophorylated derivative 2 paralleled those of 1, indicating like conformations for the two compounds. Analysis of rotating frame Overhauser spectroscopy data, coupling constants, amide proton temperature dependence, and amide proton exchange rates generated a set of constraints that were employed in energy minimization and molecular dynamics calculations using the CHARMM force field. The trans isomer exists with the tyrosine and C-terminal Tyr(+3) (Met) residues at opposite corners of the 18-membered ring separated by a distance of 16-18 A, in contrast with the cis isomer where the side chains of these residues are much closer in space (7-14 A). It was previously shown that the pTyr and the third amino acid C-terminal to this residue are the critical recognition elements for pTyr-peptide binding to the PI-3 kinase SH2 domain. Such cyclic structures may offer appropriate scaffolding for positioning important amino acid side chains of pTyr-containing peptides as a means of increasing their binding affinities to SH2 domains, and in turn provide a conceptual approach toward the design of SH2 domain directed peptidomimetics. 相似文献
959.
TR Walsh WA Neville MH Haran D Tolson DJ Payne JH Bateson AP MacGowan PM Bennett 《Canadian Metallurgical Quarterly》1998,42(2):436-439
The Aeromonas veronii bv. sobria metallo-beta-lactamase gene, imiS, was cloned. The imiS open reading frame extends for 762 bp and encodes a protein of 254 amino acids with a secreted modified protein of 227 amino acids and a predicted pI of 8.1. To confirm the predicted sequence, purified ImiS was digested and the resulting peptides were identified, yielding an identical sequence for ImiS, with 98% identity to CphA. Both possessed the putative active-site sequence Asn-Tyr-His-Thr-Asp at positions 88 to 92, which is unique to the Aeromonas metallo-beta-lactamases. 相似文献
960.
Sexual selection has been portrayed as acting predominantly on males who compete with each other over copulatory access to females; selection was considered to be driven by females choosing between males at the pre- or postcopulatory level. However, a broader view of sexual selection is now emerging. Examining male discrimination between females and female-female competition has been beneficial in identifying factors influencing the direction and strength of sexual selection. Furthermore, consideration of processes such as sexual coercion or genetic incompatibility, which indirectly influence an individual's set of copulation partners, gamete set or their offspring success, has helped to clarify the ways in which sexual selection may operate. Moreover, there is increasing evidence that not all copulations translate directly to paternity and that paternity does not necessarily translate into successful offspring. Postcopulatory and postfertilization mechanisms that influence not only paternity share but offspring recruitment now require further consideration. The benefits to each sex of copulating with particular partners or with more than one partner remains an area of debate. More carefully designed studies which eliminate alternative possibilities or quantify the relative importance of different selective pressures will also benefit from considering that not all copulations function solely to inseminate or receive sperm. It is also now clear that not all individuals of one sex follow the same strategy. Examining the variation between individuals in reproductive behaviour, fertilization success and offspring success will be important in establishing the selective pressures and mechanisms underlying the operation of sexual selection. (c) 1998 The Association for the Study of Animal Behaviour. 相似文献