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51.
Summary Changes in the intracellular distribution of microtubules and microfilaments during amoeba-to-flagellate and flagellate-to-amoeba transformations inPhysarum polycephalum were examined by fluorescence microscopy using anti-tubulin antibody and NBD-phallacidin, respectively. Amoebae contained an extensive microtubular cytoskeleton, which was converted to a flagellar cone structure during transformation to flagellates in liquid medium. When flagellates reverted back to amoebae, this conical structure disintegrated prior to flagella resorption. Amoebae showed some microfilament-enriched domains along the periphery, from which numerous filamentous extrusions, probably pseudopods and filopods, emanated. Flagellates contained a ridge, a sheet-like structure, along their dorsal axis, especially in the earlier stages of flagellation. Another microfilament-enriched thick filamentous structure ran along the dorsal axis, starting from the anterior tip of the cell. This structure apparently coincided spatially with one of the bundles of microtubules. During the reversion to amoebae, other localized microfilaments were transiently observed at the posterior end. A model of cytoskeletal changes in the transformations between these two cell types was proposed. 相似文献
52.
Peter J. M. van den Elzen Jeffrey Townsend Kathleen Y. Lee John R. Bedbrook 《Plant molecular biology》1985,5(5):299-302
Summary We show here that plant cells are sensitive to the antibiotic hygromycin-B4. We also show that a chimaeric gene consisting of the nopaline synthase (nos) gene regulatory elements and the E. coli derived hygromycin phosphotransferase (hpt) gene, when transferred to plants' cells, confers resistance to hygromycin B. The chimaeric nos-hpt gene enables efficient selection of DNA transfer to plant cells when used in conjunction with Ti plasmid-derived binary vectors in cocultivation experiments. 相似文献
53.
Christina Scharnhorst Hartmut Heinze Gabriele Meyer Waldemar Kolanus Klaus Bartsch Susanne Heinrichs Thomas Gudschun Margret Möller Frank Herzfeld 《Plant molecular biology》1985,4(4):241-245
Summary cDNA clones were isolated for a chloroplast protein, the mRNA of which is induced to maximum levels within 2–4 h after onset
of illumination in five day old, etiolated pea seedlings.
The cDNA library was constructed from poly(A)+-mRNA which was isolated from 4 h illuminated seedlings. The extremely short induction period of the early light induced protein(ELIP)-mRNA
established the basis of our screening procedure. Colony hybridization experiments were performed with32P-labelled cDNA probes, synthesized from RNA of seedlings which had been exposed to different programs of illumination. Plasmid
DNAs were isolated from colonies showing strong hybridization signals exclusively with cDNA corresponding to the 4 h-mRNA.
Hybrid released translation of preselected plasmids p 17/C2 and p17/C4 revealed a peptide of Mr 24 000. After posttranslational importin vitro, the processed product of Mr 17 000 appears in the chloroplast. Using these clones, the expression of the ELIP-mRNA was investigated by DOT-hybridization.
The ELIP-mRNA reaches maximum levels within 2–4 hours after onset of illumination. Our results correspond precisely to thein vivo characteristics and indicate positive identification of the sought clones. 相似文献
54.
S Millar D C Hayward C A Read M J Browne R V Santelli F Garcia Vallejo M T Pueyo A Zaha D M Glover F J Lara 《Gene》1985,34(1):81-86
We have constructed a library of recombinant phage containing DNA from salivary gland chromosomes of Rhynchosciara americana. We have isolated phage from this library that carry sequences homologous to cDNA clones that hybridize in situ to the DNA puffs at the polytene chromosome regions C3 and C8. This has enabled us to demonstrate a 16-fold amplification of the genomic DNA sequences at these regions during DNA-puffing. At the C8 site there is a sequence element that has characteristics of 'scrambled' moderately repetitive DNA. This is located within 3 kb from the gene encoding a 1.95-kb mRNA. We have assayed restriction fragments from the two DNA puffs for Ars activity in yeast. The only strong Ars activity is associated with a part of the moderately repetitive DNA element from the C8 puff which is not present at this site in all animals. 相似文献
55.
Abstract By using an extension to group N streptococci of a contransformation procedure we have introduced 4 different-sized cryptic plasmids for Streptococcus lactis into the plasmid-free S. lactis IL1403. A mixture of 4 cryptic plasmids with an indicator plasmid (pHV1301) conferring erythromycin resistance was used for IL1403 protoplast transformation. Under such conditions, 41.5% of the erythromycin-resistant transformants were contransformed with one of the cryptic plasmids in addition to pHV1301. Indicator plasmid pHV1301 was later spontaneously segregated from doubly transformed cells. This protocol should be very useful for constructing lactic streptococcal strains bearing any phenotypically cryptic plasmid. 相似文献
56.
The isolation and restriction mapping of a miniplasmid from the Actinomycete Nocardia corallina 总被引:1,自引:0,他引:1
Abstract A variety of plasmids has been identified as covalently closed circular and linear DNA in certain Actinomycetes, such as Streptomyces . This paper describes the first isolation and characterisation of a plasmid from the genus Nocardia . The plasmid pKU100 isolated from Nocardia corallina is a cccDNA molecule, 2.7 kb in length. This plasmid has been mapped with a wide variety of restriction enzymes and contains a number of unique restriction sites making it suitable for development as a cloning vector. 相似文献
57.
Kaoru Miyazaki Keisuke Mashima Nobuhiko Yamashita Jinpei Yamashita Takekazu Horio 《In vitro cellular & developmental biology. Plant》1985,21(1):62-66
Summary We have previously reported the transformation by Rous sarcoma virus of a cloned epithelial cell line (BRL) established from
Buffalo rat liver by H. Coon. The nontransformed (BRL) and transformed (RSV-BRL) cells grew at comparable rates in culture,
whereas only the transformed cells were tumorigenic in vivo. We report here on the existence in rat and mouse sera of a growth
inhibitor for the nontransformed BRL cells. The transformed BRL cells (RSV-BRL) were insensitive to this inhibitor. The inhibitory
activity was not prominent in sera from other species of animals tested except for rabbit; this serum inhibited the growth
of RSV-BRL cells more strongly than that of BRL cells. The growth inhibitor was partially purified from rat serum. It is a
protein free of lipid and has a molecular weight of about 220 000. The inhibitor could be separated into three components
of pI 4.6, 5.2 (major) and 5.6 by isoelectric electrophoresis.
EDITOR'S STATEMENT Although compelling theoretical arguments sometimes can be made for the likely existence of growth-inhibitory
substances of physical relevance in the control of cell proliferation, experiments aimed at identifying and studying such
factors often are difficult to design and interpret, and little strong data exists to suggest that growth-inhibitory substances
are important regulatorsin vivo. The information presented in this paper represents a start toward developing a useful system for studying growth-inhibitory
factor. David W. Barnes 相似文献
58.
Summary Studies on intercalary regeneration in several organisms have shown that a regenerate is formed when surfaces of different positional value along the proximo-distal axis are opposed. One of the main problems posed by this phenomenon is to know which piece contributes to the building of the regenerate. In the present work we have studied this problem in planarians using chimaeras made between pieces of different body levels, irradiated or not, of the sexual and asexual races ofDugesia(S)mediterranea that differ in a chromosomal marker.The results found show very clearly that intercalary regenerates in planarians are formed by cells coming from both pieces (stumps), and that irradiated pieces keep the positional values and interact with non-irradiated pieces to restore the missing parts. This means that distal and proximal transformation do actually occur at the same time during intercalary regeneration in planarians. The implications of these results as regards to the origin of cells in the regenerate and to present models of intercalary regeneration are discussed. 相似文献
59.
Summary Expression of the anchorage-independent transformed phenotype in BHK 21/13 cells generally behaves as a recessive trait. When
chemically induced and spontaneously arising transformants are fused to the nontransformed parent line, transformation is
initially suppressed, reappearing after extended growth of the hybrids. In this paper, complementation for the expression
of anchorage independence was sought among a large group of such transformants, all independently derived from BHK 21/13 cells.
Tumorigenicity studies on selected hybrids and parental lines indicated that the in vitro trait of anchorage independence
is an accurate indicator of in vivo neoplasia for these cells. Seventeen of the 18 clones tested did not complement one or
more of three tester strains. This result indicates that anchorage independence arose in these clones as a result of lesions
in the same genetic function and suggests that the final step in the progressive changes of carcinogenesis may frequently
be restricted to lesions at a single locus.
This investigation was supported by National Institutes of Health grant CA27306. 相似文献
60.
Robert T. Przygoda K. Takayama Karl A. Traul A. Tummey 《In vitro cellular & developmental biology. Plant》1985,21(1):32-38
Summary An atmosphere containing 10% CO2 has been generally accepted as optimal for the growth of Syrian hamster embryo cells in a clonal transformation assay. Data
presented in this paper show that 10% CO2 may not be the optimum environment for this assay.
Using 10 or 20% (analytically measured) CO2 in air (1 atm pressure), hamster embryo cell pools were examined for clonal growth characteristics and transformability using
five known carcinogens and a single noncarcinogenic compound. At 10% CO2, only 2 of 11 pools weee transformed by the five carcinogens but not by the noncarcinogen. At 20% CO2, six of seven pools were transformed by the five carcinogens and not by the noncarcinogen. Further, the transformation frequencies
were found to be greater in cultures incubated in an atmosphere consisting of 20% CO2 in air. The data also show that 20% CO2 increased the cloning efficiency of these cells.
A comparison of the 10 and 20% CO2 data to results reported from other conflicting interlaboratory results with this assay system may be due, in part, to variations
of CO2 concentrations. In some instances, the CO2 levels indicated by incubator flow meters vary considerably from analytically determined CO2 values. To prevent these CO2 discrepancies and their resultant effects on transformation and cloning efficiency, methods for monitoring the CO2 environment other than flow meters are recommended.
The observation of increased cloning efficiencies and transformation rates strongly suggests that culture incubation at 20%
CO2 is a preferred environment for the conduct of this assay. 相似文献