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61.
Isolation and characterization of a human interleukin 2 gene 总被引:1,自引:0,他引:1
S Mita S Maeda K Obaru N Nishino K Shimada T Hirano K Onoue T Ogawa H Ogawa 《Biochemical and biophysical research communications》1983,117(1):114-121
An interleukin 2 (IL-2) gene was isolated from a Charon 4A human gene library. Electron microscopic examination of 15 heteroduplexes formed between the genomic DNAs and the IL-2 cDNAs demonstrated that the size of the IL-2 gene is about 5.1 +/- 0.5 kb and that there are at least two introns in this gene. Nucleotide sequence of the 5' flanking region of the IL-2 gene showed a homology with that of the corresponding region of the human immune interferon gene. 相似文献
62.
Anna Maria Tagliasacchi Laura Maria Costantina Forino Manuela Frediani Silvana Avanzi 《Protoplasma》1983,115(2-3):95-103
Summary The pattern of DNA and RNA puffs in pair VII of polytene chromosomes has been investigated in the suspensor ofPhaseolus coccineus during early embryo development. The pattern of3H-TdR and3H-U incorporation has been also detected. Collected data indicate that: 1. both heterochromatic regions, p11 and q(111+112), of chromosome pair VII, organize large DNA puffs; 2. DNA puffs of both regions are specific of different embryo differentiation steps; 3. a seasonal influence on the DNA puffing seems also to be present, as demonstrated by the comparison of the results collected in two different crops; 4. the incorporation experiment by3H-TdR evidences that not all DNA puffs show clustered labeling; 5. the RNA puffing of the two regions seems also to be specific of determined embryo stages. 相似文献
63.
A simple method for electrophoretic elution of nucleic acids from gel slices is described. The procedure utilizes a standard tube gel system and can be completed in as little as one hour. Nucleic acids are recovered in a small volume with almost 100% efficiency. The procedure is applicable equally to acrylamide and agarose gels, and small as well as large RNA and DNA molecules. The eluted nucleic acids are essentially undegraded and are suitable for a variety of structural and biological analyses. 相似文献
64.
G. Stauder H. Riesemann W.M. Joester K.E. Joester 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1983,741(3)
A third DNA polymerase ‘C’ with low molecular weight was isolated and purified 3700-fold from ground hyphae of Neurospora crassa WT 74 A, which shows similarities to β- and γ-polymerases from higher eukaryotes: preference for poly(rA)(dT) as a template/primer, inhibition by p-chloromercuribenzoate, resistance against N-ethylmaleimide up to 10 mmol/l, and molecular weight of about 40 000. This polymerase elutes as a distinct peak from DEAE-cellulose at 0.60 mol/l KCl and has an optimum for K+ at 2–20 mmol/l, for Mn2+ at 0.8 mmol/l, for Mg2+ at 4.0 mmol/l, the pH optimum is 8.0. Its Km is 1.5 μmol/l using dTTP as substrate. The enzyme activity described here is free of endonuclease but contains detectable amounts of exonuclease. 相似文献
65.
T. N. DARLING D. G. DAVIS R. E. LONDON J. J. BLUM 《The Journal of eukaryotic microbiology》1989,36(2):217-225
13C-nuciear magnetic resonance (NMR) spectroscopy was used to investigate the products of glycerol and acetate metabolism released by Leishmania braziliensis panamensis promastigotes and also to examine the interaction of each of these substrates with glucose or alanine. The NMR data were supplemented by measurements of the rates of oxygen consumption and substrate utilization, and of 14CO2 production from 14C-labeIed substrate. Cells incubated with [2-13C]glycerol released acetate, succinate and D-lactate in addition to CO2. Cells incubated with acetate released only CO2. More succinate C-2/C-3 than C-l/C-4 was released from both [2-13C]glycerol and [2-13C]glucose, indicating that succinate was formed predominantly by CO2 fixation followed by reverse flux through part of the Krebs cycle. Some redistribution of the position of labeling was also seen in alanine and pyruvate, suggesting cycling through pyruvate/oxaloacetate/phosphoenolpyruvate. Cells incubated with combinations of 2 substrates consumed oxygen at the same rate as cells incubated with 1 or no substrate, even though the total substrate utilization had increased. When promastigotes were incubated with both glycerol and glucose, the rate of glucose consumption was unchanged but glycerol consumption decreased about 50%, and the rate of 14CO2 production from [l,(3)-14C]glycerol decreased about 60%. Alanine did not affect the rates of consumption of glucose or glycerol, but decreased 14CO2 production from these substrates by increasing flow of label into alanine. Although glucose decreased alanine consumption by 70%, it increased the rate of 14CO2 production from [U-14C]- and [l-14C]alanine by about 20%. This is consistent with rapid equilibration of alanine with pyruvate derived from glucose and yet little decrease in the specific activity of the large alanine pool. 相似文献
66.
RICHARD A. ALBACH 《The Journal of eukaryotic microbiology》1989,36(2):197-205
This review will concentrate on certain aspects of the nucleic acids of Entamoeba histolytica. Utilization and synthesis of purines and pyrimidines will initially be briefly discussed, e.g. salvage vs. de novo pathways, uptake studies and recognition of at least 4 transport loci. Data will be presented which show that the distribution and synthesis of RNA (to a lesser extent DNA) in the nucleus is basically the opposite one finds in other eukaryotes, viz. most RNA (ribosomal?) is synthesized (or accumulates) in the peripheral chromatin (functional equivalent of nucleolus?). The DNA is distributed and synthesized primarily throughout the nucleus. It is usually so dispersed that it will not stain with e.g. the standard Feulgen technique, unless the DNA condenses around the endosome (not a nucleolar equivalent) prior to nuclear division. Isolation of rRNA was difficult due, in part, to potent and difficult to inhibit RNase(s), some of which are apparently intimately bound to ribosomal subunits. The 25S (1.3 kDa), 178 (0.8 kDa) and 58 rRNA were recovered after isolation with a high salt SDS-DEP technique. This is the only procedure which enables us to obtain high yields of 258 rRNA: guanidine or guanidinium which permits isolation of intact functional mRNA results in isolation of small amounts of 28 RNA relative to 178 RNA. The 258 RNA is “nicked” (apparently during nuclear processing) and dissociates readily into 1 78 (0.7 kDa) and 168 (0.6 kDa) species, and a more rigidly bound 5.88 species. A small amount of “unnicked” 258 RNA was recovered with guanidine. Two DNA-dcpendent RNA polymerases (I and II) with a pronounced preference for denatured DNA as template were eluted from DEAE-Sephadex in reverse order of what occurs in other eukaryotes, except Physarum polycephalum. This conclusion was based on salt optima and alpha-amanitin sensitivity studies. Initial characterization of DNA isolated with a procedure capable of isolating > 100-kbp Leishmania DNA showed that undigested DNA migrates as a broad band between markers 6 and 24 kbp. The persistent recovery of such a “band” by us and Perez-Mutul et al. no larger than ca. 24 kbp (with the exception of >48 kbp DNA isolated by Hernandez et al. using an in situ lysis technique which did not include a proteinase) may be due to nicks introduced during isolation; or, perhaps much of the amebal DNA exists in vivo as gene sized fragments. However, preliminary data generated using orthogonal pulse-field agarose gel electrophoresis do suggest that amebal DNA may be in small chromosomes. 相似文献
67.
Tsuneyoshi Kuroiwa 《Journal of plant research》1989,102(2):291-329
It has been established that organelles, such as mitochondria and plastids, contain organelle-specific DNA and arise from
the division of pre-existing organelles (e.g., Possingham and Lawrence, 1983). We propose that organelle DNAs, such as mitochondrial
DNA and plastid DNA are not naked in organellesin situ but are organized in each case to form an “organelle nucleus” with basic proteins (Kuroiwa, 1982). The concept of organelle
nuclei has changed our ideas about the division of organelles. Thus, the process of organelle division must be composed of
two main events: division of the organelle nucleus and organellekinesis (division of the other components of the mitochondrion
or plastid). The latter term has been adopted as an appropriate analogue of cytokinesis.
We were the first to identify the plastid-dividing ring (PD-ring), which is located in the cytoplasm close to the outer envelope
membrane at the constricted isthmus of dividing chloroplasts in the red algaCyanidium caldirum. The PD-ring is about 60 nm in width and 25 nm in thickness, and is a circular bundle of actin-like, fine filaments, each
about 4–5 nm in diameter. Since cytochalasin B, an inhibitor of polymerization of actin filaments, inhibits the formation
of the PD-ring and, thus, prevents subsequent division of chloroplasts, the PD-ring is thought to be a structure that is essential
for the division of plastids (plastidkinesis).
The behavior of the PD-ring during a cycle of chloroplast division can be classified into the following four stages on the
basis of morphological and temporal differences. The chloroplast growth stage: the small, spherical chloroplast increases
in volume and becomes a football-like structure, while the PD-ring from the previous division disappears. Formation of the
PD-ring: the somewhat electron-dense body (see below) is fragmented into many, somewhat electron-dense granules, which are
aligned along the equatorial region of the chloroplast and fine filaments are formed from the somewhat electron-dense granules
in the equatorial region. The fine filaments of the PD-ring align themselves according to the longest axis of their overall
domain, i.e., circumferentially. Contraction stage: a bundle of fine filaments begins to contract and generates a deep furrow.
Conversion stage: after chloroplast division, the remnants of the PD-ring are converted into somewhat electron-dense bodies.
Similar events occur during the second cycle of chloroplast division. Since similar structures are observed extensively in
the plastids of algae, moss and higher plants, the PD-ring appears to be an essential structure for the division of plastids
in plants. 相似文献
68.
Bradley C. Hyman 《Journal of nematology》1990,22(1):24-30
Genetic variation within nuclear and mitochondrial DNA of Meloidogyne species and host races has been evaluated for the development of root-knot nematode molecular diagnostics. This review summarizes the distinctive features of several useful DNA-based assays for plant-parasitic nematodes, focusing upon the direct application of these procedures for Meloidogyne detection, identification, and systematics. 相似文献
69.
I. Negrutiu J. Dewulf M. Pietrzak J. Botterman E. Rietveld E. M. Wurzer-Figurelli De Ye M. Jacobs 《Physiologia plantarum》1990,79(1):197-205
Reports on direct gene transfer have dealt with either the obtention of stable transformants and transgenic plants, or described the use of reporter genes to analyse different aspects of gene expression in plant protoplasts and conditions for their use in transient gene expression assays.
In this paper we present comparisons between several transformation techniques, show species-specific differences in efficiencies of stable transformants and in the levels of transient gene expression, and report on the identification of major parameters responsible for DNA uptake as judged from transient chloramphenicol acetyl transferase (CAT) expression levels and from efficiencies of transformation based on kanamycin-resistance. The described procedures have been simplified, optimized and standardized and should allow routine use with a great variety of plant species. 相似文献
In this paper we present comparisons between several transformation techniques, show species-specific differences in efficiencies of stable transformants and in the levels of transient gene expression, and report on the identification of major parameters responsible for DNA uptake as judged from transient chloramphenicol acetyl transferase (CAT) expression levels and from efficiencies of transformation based on kanamycin-resistance. The described procedures have been simplified, optimized and standardized and should allow routine use with a great variety of plant species. 相似文献
70.
Thymocyte growth peptide (TGP) initiates DNA synthesis in immature thymocytes and has previously been characterized as an acidic peptide isolated from calf thymus. We now report the isolation of TGP from sheep thymus and show it to be a nonapeptide with a large N-terminal blocking moiety characterized by high UV absorbance. The amino acid composition is identical to FTS, consisting of 2 Gly, 2 Ser, 2 Glx, 1 Ala, 1 Lys, 1 Asx. In contrast to FTS, TGP is acidic with an apparent isoelectric point of 4.2 and a high UV absorbance at 270–280 nm. Reverse phase chromatography of TGP at an acidic pH results in a change of the molecule and the appearance of two new compounds TGP-A and TGP-B, both with less than 50% of the original TGP activity. Full activity could be restored by the addition of ZnCl2 to TGP-A. Both TGP-A and B have some amino acid composition and high UV absorbance as native TGP. We propose that TGP consists of a non-peptide moiety bound to the N-terminal of the nonapeptide Glu-Ala-Lys-Ser-Gln-Gly-Gly-Ser-Asn and that the active molecule is stabilized by Zn2+. 相似文献