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51.
Cells of Escherichia coli induced for l-tryptophan synthase [l-serine hydro-lyase (adding indole-glycerol-phosphate), EC 4.2.1.20] have been assayed in DMF and DMSO aqueous solvents as reaction medium. Up to 20% DMF/water, cells retained 90% of their tryptophan synthase activity. Concentrations of 20 mM indole, which did not inhibit this reactivity, could be reached with 5% DMF/water. Four matrices were compared for cell immobilization: polyacrylamide, foam particles of bovine seum albumin, alginate and κ-carrageenan. The best activity was retained with the latter matrix, and the preparations thus obtained allowed high productivity of l-tryptophan. Various systems of production of l-tryptophan with κ-carrageenan and DMF/water were studied. 相似文献
52.
Summary
Claviceps purpurea strain 129 was cultivated under submerged conditions in a sucrose-citrate medium containing high (36.8 mM) or low (1.84 mM) KH2PO4 concentrations. The permeabilized cells and culture supernatants contained alkaline and acid phosphatases. In the medium containing a high phosphate concentration, the synthesis of extracellular phosphatases was repressed, but that of cellular phosphatases was not. Extracellular phosphatases, especially alkaline phosphatases, were derepressed by transferring the mycelium into a phosphate-free medium. This derepression was inhibited by cycloheximide. In the presence of cycloheximide, the activities of the cellular phosphatases decreased markedly, indicating turnover of these enzymes. The cellular acid phosphatase was inhibited by phosphate (0.025 M–0.1 M) and NaF (0.01 M) while the cellular alkaline phosphatase was only inhibited by phosphate. Both cellular and extracellular alkaline phosphatases were more sensitive to repression by phosphate than the acid phosphatases. The alkaloid synthesizing enzymes were: a) present in mycelia grown in high levels of phosphate and b) activated by decreasing the intracellular phosphate level. 相似文献
53.
54.
Debaryomyces phaffii inulinase was immobilized by adsorption on DEAE-cellulose. The immobilization caused a drop in optimum pH, a slight increase in optimum temperature and an important increase in the thermal stability of the system. The activity yield of immobilized inulinase was 75%.A continuous reactor operation was carried out. The utilization of this system permited the production of fructose syrup from inulin. 相似文献
55.
Dominique Weil Nguyen Van-Cong Catherine Finaz R. Rebourcet Chantal Cochet J. de Grouchy J. Frézal 《Human genetics》1977,36(2):205-211
Summary 22 independent man-hamster (HGPRT–) hybrids using male human cells with balanced reciprocal translocation t(X;2)(p22;q32) were analysed for human genes localized on chromosome 2 (IDHS, MDHS), on chromosome X (PGK, GAL, G6PD) and for the different chromosomes in relation with the balanced reciprocal translocation (chr.2, chr.2q–, chr.Xp+).The following results were obtained:The chromosomes 2 and 2q– are absent in the 22 hybrids.In 9 hybrids, the absence of MDHS in spite of the presence of the chromosome Xp+ indicates that the gene for MDHS is not localized on this chromosome (or that the gene for MDHS is not on the segment 2q32-2qter translocated on X).In 14 hybrids, the three markers of X (PGK, GAL, G6PD) and IDHS are expressed in the presence of the chromosome Xp+. This result indicates that the genes for these markers are on Xp+ or that the genes PGK, GAL, G6PD are on X without the Xp22-Xter segment, translocated on the chr.2, and that the gene for IDHS is on the 2q32-2qter segment translocated on X.In 8 hybrids, in the absence of the intact chromosome Xp+, the higher percentage of the presence of G6PD (7 hybrids) and the lower percentage of the presence of IDHS (3 hybrids) are explained by the fact that these hybrids selected in HAT medium had to retain a segment of Xp+ bearing the human gene HGPRT. G6PD appeared very close to HGPRT and IDHS very distant from HGPRT.The study of the different correlations between the presence and the absence of these four markers on Xp+ in the different hybrids indicates the following order on the chromosome Xp+ from p to q: IDHs — PGK — GAL — G6PD.
Groupe INSERM: Directeur J. Frézal
Groupe CNRS, ER, 149: Directeur J. de Grouchy 相似文献
Groupe INSERM: Directeur J. Frézal
Groupe CNRS, ER, 149: Directeur J. de Grouchy 相似文献
56.
A.C. Garapin B. Cami W. Roskam P. Kourilsky J.P. Le Pennec F. Perrin P. Gerlinger M. Cochet P. Chambon 《Cell》1978,14(3):629-639
The Eco RI fragment “b” of chicken DNA (Breathnach, Mandel and Chambon, 1977), which contains the sequences coding for the 5′ quarter of ovalbumin mRNA (ov mRNA), has been isolated by molecular cloning using a “shotgun” approach. Electron microscopy and restriction enzyme analysis have revealed that the sequences coding for the 5′ quarter (~500 nucleotides) of ov mRNA are split into four regions separated by three intervening sequences. The cloning procedure seems to be reliable, since the restriction enzyme pattern of the cloned Eco RI fragment “b” is similar to that of the corresponding chromosomal DNA fragment. There is no evidence supporting the existence of a 150–200 nucleotide long sequence at the 5′ end of the ov mRNA similar to the “leader” sequences found at the 5′ end of some adenovirus and SV40 mRNAs. 相似文献
57.
Alfred Berteloot Christiane Malo Sylvie Breton Michel Brunette 《The Journal of membrane biology》1991,122(2):111-125
Summary Kinetic data in (brush-border) membrane vesicles which rely on the validity of the initial rate assumption for their interpretation and depend on tracer flux studies using the rapid filtration technique for their experimental measurement have been limited to some extent by the absence of techniques that would allow for real-time data analysis. In this paper, we report on our successful design of a fast sampling, rapid filtration apparatus (FSRFA) which seems to fill up this technical gap since showing the following characteristics: (i) rapid injection (5 msec) and mixing (less than 100 msec) of small amounts of vesicles (10–40 l) with an incubation medium (0.2–1.0 ml); (ii) fast (20 to 80 msec depending on the sample volume) and multiple (up to 18 samples at a maximal rate of 4/sec) sampling of the uptake mixture followed by rapid quenching in the stop solution (approximately 5 msec) according to a predetermined time schedule (any time combination from 0.25 to 9999 sec); and (iii) fast, automated, and sampling-synchronized filtration and washings of the quenched uptake medium (only 15–20 sec are necessary for the first filtration followed by two washings and extra filtrations). As demonstrated using adult human jejunal brush-border membrane vesicles and Na+-d-glucose cotransport as models, the FSRFA accurately reproduces the manual aspects of the rapid filtration technique while allowing for very precise initial rate determinations. Moreover, the FSRFA has also been designed to provide as much versatility as possible and, in its present version, allows for a very precise control of the incubation temperature and also permits a few efflux protocols to be performed. Finally, its modular design, which separates the fast sampling unit from the rapid filtration device, should help in extending its use to fields other than transport measurement. 相似文献
58.
Armelle Baeza-Squiban Sylvie Romet Anne Moreau Francelyne Marano 《In vitro cellular & developmental biology. Animal》1991,27(6):453-460
Summary Primary cultures of rabbit tracheal cells were obtained as outgrowths from explants of tracheal mucosa. A 30% collagen substratum
containing serum and minimal essential medium was required for obtaining an outgrowth of epithelial cells keeping their differentiated
characteristics. The tracheal epithelial cells obtained near the explant in the first days of culture presented morphologic
similarities with normal tracheal epithelium. Cultures contained basal cells and epithelial polarized cells that exhibited
apical tight junctions and desmosomes. Ciliated cells stayed functional during all time culture. Their number slightly increased
at the beginning of the culture and then stayed constant when the total number of cells increased. Development of the outgrowth
was rapid and significant inasmuch as the outgrowth surface reached 30 times that of the explant after less than 8 days. This
was linked to cellular proliferation, as demonstrated by the incorporation of bromodeoxyuridine (BrdU) in phase-S nuclei and
the revelation of BrdU using an immunofluorescence technique. The epithelial nature of the outgrowth cells and the absence
of contamination with fibroblasts were established by positive staining with anti-keratin antibody and by negative staining
with anti-vimentin antibody, respectively.
This work was supported by DRET and by CIFRE grant awarded to S. R. 相似文献
59.
Sylvie Tutois Josselyne Salaun Marie-Geneviève Mattei Jean-Louis Guénet 《Mammalian genome》1991,1(3):184-190
Transgenic mice generated with different DNA sequences were surveyed for possible homozygous mutant phenotypes. We found an embryonic lethal mutation in the transgenic mouse strain (MT-MYC12.4) containing the human c-myc gene. Embryos homozygous for the transgene die shortly after implantation. The strain MT-MYC12.4 carries approximately 50 tandem copies of the recombinant plasmid sequence. The 3 flanking sequence has been cloned and analyzed. It contains a unique sequence that has been conserved during evolution and maps to Chromosome (Chr) 9. This mutant has been designated Tg 9 (HSA-MYC). 相似文献
60.