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1.
K. H. Jang J. W. Seo K. B. Song C. H. Kim S. K. Rhee 《Bioprocess and biosystems engineering》1999,21(5):453-458
Secretion of levansucrase from Zymomonas mobilis in Escherichiacoli by glycine supplement was investigated. A significant amount of levansucrase (about 25% of total activity) was found in intact whole-cells. Cell fractionation experiments showed that levansucrase was found both in the periplasmic space and in the cytoplasmic fraction of E. coli. None or only trace amounts of levansucrase was detected in the extracellular culture broth at 24 h of cultivation and it accrued with the increasing concentration of glycine in the culture medium and duration of the culture period. Optimal glycine concentration for the maximum secretion of levansucrase was in the range of 0.8-1%, in which approximately 20-50% of levansucrase was released into the extracellular fraction at 24 h of cultivation, although glycine retarded the bacterial growth. 相似文献
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Cascade control of Escherichia coli glutamine synthetase. Purification and properties of PII protein and nucleotide sequence of its structural gene 总被引:20,自引:0,他引:20
A procedure was developed to purify large quantities of PII protein from an Escherichia coli strain which contains a multicopy plasmid harboring the structural gene of PII (the glnB gene). Ultraviolet spectra of uridylylated and unuridylylated PII were obtained using the purified PII and empirical formulas to calculate the concentration of protein and the average number of uridylylated subunits per molecule were derived. A continuous fluorometric assay for the measurement of uridylylated PII (PIID) and adenylyltransferase (ATase) was also established. Rate measurements at various concentrations of PIID and at a fixed concentration of ATase showed that a tetrameric PIID molecule interacts with only one ATase molecule at a time. The complete nucleotide sequence of the glnB gene was determined and parts of the deduced amino acid sequence were confirmed by the results of amino acid sequence analysis of peptides. The PII subunit consists of 103 amino acids (Mr = 11,580). Two tyrosines reside at positions 46 and 51, where Tyr51 is the site of uridylylation. Nucleotide sequence analysis of the upstream region showed no obvious sites for the binding of RNA polymerase, indicating that the glnB gene is a part of an as yet unidentified operon. 相似文献
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A Rapid Screening Test for Aflatoxin-synthesizing Aspergilli of the flavus-oryzae Group 总被引:1,自引:1,他引:0
P. de Vogel Renée van Rhee W. A. A. Blanche Koelensmid 《Journal of applied microbiology》1965,28(2):213-220
A rapid test for the recognition of aflatoxin-synthesizing strains of the Aspergillus flavus–oryzae group is described. For this purpose the strains are cultivated on Czapek–Dox agar enriched with an aqueous extract of groundnuts, and in which sodium nitrate is replaced by ammonium chloride. Toxin production is observed by the production of a bright blue fluorescence in the medium when placed under an ultraviolet lamp. 相似文献
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SPA::EcoRI fusion protein was produced by Escherichia coli JM103 carrying the multicopy expression plasmid pMTC48, the multicopy repressor plasmid pRK248, and the multicopy protection plasmid pEcoR4 in a 60-L working volume airlift tower loop reactor on M9 minimal medium with glucose. Cell mass concentration, total cell count, number of colony-forming units, specific growth rate, yield coefficient, and metabolite (acetate, pyruvate, succinate, lactate, ethanol) concentrations were monitored during the growth phase and gene expression. Gene expression was induced by temperature shift or chemically by isopropyl-thiogalactosidase in the airlift tower loop reactor (ALTR) at constant cultivation time and in a small stirred tank reactor at different cultivation times. During induction, the cultivation medium was supplemented with concentrated Luria-Bertani (LB) medium. The intracellular enzyme activity was evaluated as a function of the time after the start of the induction. It was found that the reduction of the glucose concentration and increase of the dissolved oxygen concentration reduced the acetate produced and increased the intracellular enzyme activity. (c) 1993 John Wiley & Sons, Inc. 相似文献
8.
S G Rhee J J Villafranca P B Chock E R Stadtman 《Biochemical and biophysical research communications》1977,78(1):244-250
Glutamine synthetase from is modulated by adenylylation of a tyrosine residue on each subunit of the dodecamer, as well as by feedback inhibition. With the stopped-flow fluorometric method, the binding constants for L-Glu, L-Ala, D-Val, and Gly to E1.0—Mg, E7, in the absence or presence of ATP or ADP, and NH3 were evaluated at pH 7.0, 15°. Strong synergistic effects between the amino acids and the nucleotide were observed. The fluorescence amplitude observed due to either simultaneous or sequential addition of 2 different amino acids to E or E·ATP indicate that L-Glu can bind to the enzyme simultaneously with L-Ala, Gly and D-Val; L-Ala can coexist with D-Val, Gly or D-Ala. NMR method also shows that L-Glu and L-Ala can bind simultaneously. Therefore, within our experimental conditions, the unadenylylated enzyme possesses allosteric site(s) for the amino acid inhibitors. 相似文献
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Anabaena sp., isolated from a rice paddy, was investigated for its nitrogen fixation as measured by acetylene reduction activity (ARA)
in P-limited continuous and light-limited semi-continuous cultures. Growth rate (μ) under P limitation was a function of cell
P content (q
p). Both the photosynthetic capacity (Pmax) and photosynthetic efficiency (α) increased with μ when expressed per cell, but not per unit chla. The ARA of steady-state cells under P limitation increased with μ and was linearly related to C-fixation rate. This was
apparently a consequence of the control of C-fixation by P limitation. In light-limited cells, steady state ARA, both at the
culture light intensity and in the dark, increased asymptotically with μ, but the activity in the dark was only about 51%
of that in the light. When the light level of steady-state cells grown at a high in intensity was switched to a low level,
ARA decreased exponentially with time. Dark ARA activity also showed a similar decline, but at much lower levels. Thus, ARA
depended not only on light history, but also immediate photosynthesis. Steady-state ARA at the ambient intensity or in the
dark showed a strong correlation with14C-fixation rate. ARA of light-limited cells showed the same light-saturation characteristics as their14C-fixation, with the same initial saturation intensity,I
k. The ratios of Pmax to the maximum ARA (ARAmax), and α to the slope of ARA (αara) were identical. A comparison of gross to net photosynthesis and N2 fixation suggested that there was little leakage or excretion of fixed C or N. 相似文献