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21.
The incubation of peptidoglycan fragments with ether-treated cells of Neisseria gonorrhoeae resulted in breakdown products that showed the presence of previously undescribed lytic enzymes. The properties of an endopeptidase able to hydrolyse peptide-linked bis-disaccharide peptide dimer to monomer units were examined. An exo-N-acetyl-glucosaminidase was also shown to release free N-acetylglucosamine. The breakdown pattern of glycosidically-linked dimer indicated the existence of an endo-N-acetylglucosaminidase. The activities of the latter enzyme and of the endopeptidase were both sensitive to beta-lactam antibiotics. 相似文献
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R E Perkins S C Conroy B Dunbar L A Fothergill M F Tuite M J Dobson S M Kingsman A J Kingsman 《The Biochemical journal》1983,211(1):199-218
The complete amino acid sequence of yeast phosphoglycerate kinase, comprising 415 residues, was determined. The sequence of residues 1-173 was deduced mainly from nucleotide sequence analysis of a series of overlapping fragments derived from the relevant portion of a 2.95-kilobase endonuclease-HindIII-digest fragment containing the yeast phosphoglycerate kinase gene. The sequence of residues 174-415 was deduced mainly from amino acid sequence analysis of three CNBr-cleavage fragments, and from peptides derived from these fragments after digestion by a number of proteolytic enzymes. Cleavage at the two tryptophan residues with o-iodosobenzoic acid was also used to isolate fragments suitable for amino acid sequence analysis. Determination of the complete sequence now allows a detailed interpretation of the existing high-resolution X-ray-crystallographic structure. The sequence -Ile-Ile-Gly-Gly-Gly- occurs twice in distant parts of the linear sequence (residues 232-236 and 367-371). Both these regions contribute to the nucleoside phosphate-binding site. A comparison of the sequence of yeast phosphoglycerate kinase reported here with the sequences of phosphoglycerate kinase from horse muscle and human erythrocytes shows that the yeast enzyme is 64% identical with the mammalian enzymes. The yeast has strikingly fewer methionine, cysteine and tryptophan residues. 相似文献
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Jean-Marie Frère Jean-Marie Ghuysen Peter E. Reynolds Ramon Moreno Harold R. Perkins 《The Biochemical journal》1974,143(1):241-249
Benzylpenicillin and cephaloridine reacted with the exocellular dd-carboxypeptidase–transpeptidase from Streptomyces R39 to form equimolar and inactive antibiotic–enzyme complexes. At saturation, the molar ratio of chromogenic cephalosporin 87-312 to enzyme was 1.3:1, but this discrepancy might be due to a lack of accuracy in the measurement of the antibiotic. Spectrophotometric studies showed that binding of cephaloridine and cephalosporin 87-312 to the enzyme caused opening of their β-lactam rings. Benzylpenicillin and cephalosporin 87-312 competed for the same site on the free enzyme, suggesting that binding of benzylpenicillin also resulted in the opening of its β-lactam ring. In Tris–NaCl–MgCl2 buffer at pH7.7 and 37°C, the rate constants for the dissociation of the antibiotic–enzyme complexes were 2.8×10−6, 1.5×10−6 and 0.63×10−6s−1 (half-lives 70, 130 and 300h) for benzylpenicillin, cephalosporin 87-312 and cephaloridine respectively. During the process, the protein underwent reactivation. The enzyme that was regenerated from its complex with benzylpenicillin was as sensitive to fresh benzylpenicillin as the native enzyme. With [14C]benzylpenicillin, the released radioactive compound was neither benzylpenicillin nor benzylpenicilloic acid. The Streptomyces R39 enzyme thus behaved as a β-lactam-antibiotic-destroying enzyme but did not function as a β-lactamase. Incubation at 37°C in 0.01m-phosphate buffer, pH7.0, and in the same buffer supplemented with sodium dodecyl sulphate caused a more rapid reversion of the [14C]benzylpenicillin–enzyme complex. The rate constants were 1.6×10−5s−1 and 0.8×10−4s−1 respectively. Under these conditions, however, there was no concomitant reactivation of the enzyme and the released radioactive compound(s) appeared not to be the same as before. The Streptomyces R39 enzyme and the exocellular dd-carboxypeptidase–transpeptidase from Streptomyces R61 appeared to differ from each other with regard to the topography of their penicillin-binding site. 相似文献
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E M Perkins 《American journal of physical anthropology》1968,29(3):349-364
The pigmy marmoset — Callithrix (= Cebuella) pygmaea Spix — is the second detailed study of the members of the family Callithricidae. It is closely allied to the red-mantled tamarin — Saguinus (=Tamarinus) fuscicollis illigeri Spix — and shares some of its characteristics with both Prosimii and Anthropoidea. The epidermis and dermis contain moderate numbers of concurrent, melanotic melanocytes. The dermis is rich in elastin. Hair follicles grow in groups of three or four over the general body surface, and one apocrine gland is associated with each grouping. Arrectores pilorum muscles are well developed. On the ventral ulnar wrist are sinus hairs associated with apocrine glands. Most hair follicles have nerve end-organs around them that are reactive for acetylcholinesterase and alkaline phosphatase. There is a large aggregation of sebaceous glands in the suprapubic region. The large sebaceous glands in the eyelid, face, and external genitalia are surrounded by cholinesterasereactive nerves. Apocrine glands are found over most of the hairy skin except the brow, scalp and back; a large grouping of them is present in the sternal region. Only the secretory coils of apocrine glands in the external genitalia are invested with butyrylcholinesterase-rich nerves. Eccrine glands are confined to the volar surfaces of the pes and manus. They have dark cells with abundant glycogen and clear cells with neither glycogen nor PAS-reactive material. The nerves around the eccrine secretory coil are reactive only for acetylcholinesterase. 相似文献
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