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61.
A yeast genomic library in Escherichia coli, constructed by insertion of Sau3A restriction fragments into the hybrid Saccharomyces cerevisiae-E. coli plasmid pFL1, was screened by a radioimmunoassay (RIA) for colonies expressing yeast aspartyl-tRNA synthetase (AspRS). Four clones were isolated by this technique. Data obtained by Southern and restriction analysis of the inserts showed a common 3.8-kb BamHI restriction fragment which, when inserted into the plasmid pFLl, gave a positive RIA. Several controls showed that this 3.8-kb insert codes for the entire AspRS : (i) S. cerevisiae transformed by the PFL1 plasmid carrying the 3.8-kb fragment overproduces AspRS activity by a factor of ten compared to the wild-type yeast strain; and (ii) a new protein with electrophoretic behaviour similar to AspRS and immuno-reactive toward anti-AspRS appears in crude extracts of transformed yeast and E. coli. 相似文献
62.
Dr. F. James Bailey Janet Blankenship Jon H. Condra Robert Z. Maigetter Ronald W. Ellis 《Journal of industrial microbiology & biotechnology》1987,2(1):47-52
Summary Studies are presented on the fermentation of recombinantEscherichia coli that express rat atrial natriuretic factor (ANF) as a fusion protein. Our objective was to achieve high cell density while maintaining ANF expression at the same level as observed in shake flasks. Improved fermentation conditions included: maintaining glucose concentrations at 1 g/l, using an enriched medium, adding concentrates of medium throughout the fermentation, and blending oxygen for adequate aeration. Cell densities of 12 g/l (dry weight) were achieved, which represented a 10-fold increase over non-improved conditions, while maintaining ANF levels at 7 mg/g of dry cell mass. When galactose was used as an initial carbon source or as a feed supplement, there was a 2-3-fold increase in the expression of ANF from these high-cell-density fermentations. The recombinant ANF was biologically active. 相似文献
63.
Abstract Enteropathogenic strains of faecal Escherichia coli produced significantly ( P < 0.01) more maltase than the non-pathogenic strains of the organism. The enzyme was induced by maltose but repressed by glucose and fructose. The maltase was partially purified by ammonium sulphate precipitation, followed by dialysis and gel permeation chromatography. The partially purified maltase had an M r of 144500 and an apparent K m of approx. 7.6 mM for maltose. The enzyme was stimulated by Ca2+ , inhibited by Cu2+ , Hg2+ , Uo2+ , IAA and EDTA, and exhibited optimum activity at pH 6.5 at 30°C. 相似文献
64.
Abstract The first step of aerobactin biosynthesis, oxidation of an aliphatic primary amino group to an N -hydroxy-amino compound seems to be involved in the biosynthesis of most of the hydroxamatetype siderophores which are widely distributed among bacteria and fungi. Therefore, the first step of aerobactin biosynthesis, oxidation of lysine to N 6 -hydroxylysine was studied as a model reaction using a strain of Escherichia coli that contains the first gene aerA of aerobactin synthesis on a multi-copy plasmid and which is lacking the gene for the subsequent step in the pathway. In addition, culture conditions are described which lead to the secretion of N 6 -hydroxylysine into the medium in amounts that can easily be quantitatively determined by a simple, reliable chemical assay. This assay can be used for screening inhibitors of the oxidation of α-amino groups, which should interfere with the biosynthesis of siderophore hydroxamates and thus should create bacteriostatic conditions. 相似文献
65.
Abstract Subinhibitory concentrations of trimethoprim-sulfamethoxazole increased the total yield of Shiga-like toxin (SLT), produced by Shigella dysenteria 1 and by enterophathogenic and enterohemorrhagic strains of Escherichia coli . Stimulation of SLT synthesis by trimethoprim-sulfamethoxazole was demonstrated by an increase in cytotoxic activity for HeLa cells and the diameter of the zone formed around bacterial colonies probed with monoclonal antibodies to SLT. Thus, supplementation of culture media with trimetroprimsulfamethoxazol will facilitate SLT purification and detection of SLT-producing bacteria. 相似文献
66.
Jörg Hacker Manfred Ott Günter Schmidt Richard Hull Werner Goebel 《FEMS microbiology letters》1986,36(2-3):139-144
Abstract The genetic determinant coding for the P-specific F8 fimbriae was cloned from the chromosome of the Escherichia coli wild-type strain 2980 (O18:K5:H5:F1C, F8). The F8 determinant was further subcloned into the Pst I site of pBR322 and a restriction map was established. In a Southern hybridization experiment identity between the chromosomally encoded F8 determinant of 2980 and its cloned counterpart was demonstrated. The cloned F8 fimbriae and those of the wild type strain consist of a protein subunit of nearly 20 kDa. F8 fimbriated strains were agglutinated by an F8 polyclonal antiserum, caused mannose-resistant hemagglutination and attached to human uroepithelial cells. The cloned F8 determinant was well expressed in a variety of host strains. 相似文献
67.
68.
Giorgio Brandi Flaminio Cattabeni Amedeo Albano Orazio Cantoni 《Free radical research》1989,6(1):47-55
Escherichia coli lethality by hydrogen peroxide is characterized by two modes of killing. In this paper we have found that hydroxyl radicals (OH -) generated by H2O2 and intracellular divalent iron are not involved in the induction of mode one lethality (i.e. cell killing produced by concentrations of H2O2 lower than 2.5 mM). In fact, the OH radical scavengers, thiourea, ethanol and dimethyl sulfoxide, and the iron chelator, desferrioxarnine, did not affect the survival of cells exposed to 2.5mM H2O2. In addition cell vulnerability to the same H2O2 concentration was independent on the intracellular iron content. In contrast, mode two lethality (i.e. cell killing generated by concentrations of H2O2 higher than 10mM) was markedly reduced by OH radical scavengers and desferrioxamine and was augmented by increasing the intracellular iron content.
It is concluded that OH. are required for mode two killing of E. coli by hydrogen peroxide. 相似文献
It is concluded that OH. are required for mode two killing of E. coli by hydrogen peroxide. 相似文献
69.
70.
Molecular and functional analysis of the ruv region of Escherichia coli K-12 reveals three genes involved in DNA repair and recombination 总被引:14,自引:0,他引:14
Gary J. Sharples Fiona E. Benson Graham T. Illing Robert G. Lloyd 《Molecular & general genetics : MGG》1990,221(2):219-226
Summary Recombinant plasmids carrying ruvA, ruvB, or both were constructed and used to investigate the genetic defects in a collection of UV-sensitive ruv mutants. The results revealed that efficient survival of UV-irradiated cells depends on both ruvA and ruvB, and on a third gene, ruvC, located upstream of the ruvAB operon. Southern blotting analysis was used to locate insertions in ruv and to examine putative deletion mutants. Two Tn10 insertions were located to the region encoding ruvA. Since these insertions caused a deficiency in the activities of both ruvA and ruvB, we concluded that they must exert a polar effect on ruvB. Two putative ruv deletion mutants were shown to be the result of deletion-inversion events mediated during imprecise excision of Tn10. The relevant inversion breakpoints in these mutants were located to ruvA and ruvC. 相似文献