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41.
Cloning of cDNAs for human phosphoribosylpyrophosphate synthetases 1 and 2 and X chromosome localization of PRPS1 and PRPS2 genes 总被引:3,自引:0,他引:3
M A Becker S A Heidler G I Bell S Seino M M Le Beau C A Westbrook W Neuman L J Shapiro T K Mohandas B J Roessler 《Genomics》1990,8(3):555-561
Cloned cDNAs representing the entire, homologous (80%) translated sequences of human phosphoribosylpyrophosphate synthetase (PRS) 1 and PRS 2 cDNAs were utilized as probes to localize the corresponding human PRPS1 and PRPS2 genes, previously reported to be X chromosome linked. PRPS1 and PRPS2 loci mapped to the intervals Xq22-q24 and Xp22.2-p22.3, respectively, using a combination of in situ chromosomal hybridization and human x rodent somatic cell panel genomic DNA hybridization analyses. A PRPS1-related gene or pseudogene (PRPS1L2) was also identified using in situ chromosomal hybridization at 9q33-q34. Human HPRT and PRPS1 loci are not closely linked. Despite marked cDNA and deduced amino acid sequence homology, human PRS 1 and PRS 2 isoforms are encoded by genes widely separated on the X chromosome. 相似文献
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Determinants of mRNA stability in Dictyostelium discoideum amoebae: differences in poly(A) tail length, ribosome loading, and mRNA size cannot account for the heterogeneity of mRNA decay rates. 总被引:9,自引:6,他引:3
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R A Shapiro D Herrick R E Manrow D Blinder A Jacobson 《Molecular and cellular biology》1988,8(5):1957-1969
As an approach to understanding the structures and mechanisms which determine mRNA decay rates, we have cloned and begun to characterize cDNAs which encode mRNAs representative of the stability extremes in the poly(A)+ RNA population of Dictyostelium discoideum amoebae. The cDNA clones were identified in a screening procedure which was based on the occurrence of poly(A) shortening during mRNA aging. mRNA half-lives were determined by hybridization of poly(A)+ RNA, isolated from cells labeled in a 32PO4 pulse-chase, to dots of excess cloned DNA. Individual mRNAs decayed with unique first-order decay rates ranging from 0.9 to 9.6 h, indicating that the complex decay kinetics of total poly(A)+ RNA in D. discoideum amoebae reflect the sum of the decay rates of individual mRNAs. Using specific probes derived from these cDNA clones, we have compared the sizes, extents of ribosome loading, and poly(A) tail lengths of stable, moderately stable, and unstable mRNAs. We found (i) no correlation between mRNA size and decay rate; (ii) no significant difference in the number of ribosomes per unit length of stable versus unstable mRNAs, and (iii) a general inverse relationship between mRNA decay rates and poly(A) tail lengths. Collectively, these observations indicate that mRNA decay in D. discoideum amoebae cannot be explained in terms of random nucleolytic events. The possibility that specific 3'-structural determinants can confer mRNA instability is suggested by a comparison of the labeling and turnover kinetics of different actin mRNAs. A correlation was observed between the steady-state percentage of a given mRNA found in polysomes and its degree of instability; i.e., unstable mRNAs were more efficiently recruited into polysomes than stable mRNAs. Since stable mRNAs are, on average, "older" than unstable mRNAs, this correlation may reflect a translational role for mRNA modifications that change in a time-dependent manner. Our previous studies have demonstrated both a time-dependent shortening and a possible translational role for the 3' poly(A) tracts of mRNA. We suggest, therefore, that the observed differences in the translational efficiency of stable and unstable mRNAs may, in part, be attributable to differences in steady-state poly(A) tail lengths. 相似文献
44.
Dawson K 《Bioethics》1988,2(1):1-14
A basic consideration in research on human embryos is the controversy about when the embryo acquires moral status. The author refutes the contention that segmentation is the determinant of moral status. She notes that segmentation, as a stage in embryonic development, does not coincide with the development of "irreversible individuality" upon which the segmentation argument depends. Dawson also finds a lack of clarity in the meaning of "individuality." These problems, she maintains, prevent segmentation from being morally important and render the proposed 14-day limit on embryo research unnecessary. Dawson concludes that to introduce a time restriction on embryo research is premature because it is based on an inadequate philosophical argument. 相似文献
45.
Karen Barsuhn S. Theodore Chester Jr. Dr. Jane A. Z. Leedle 《Current microbiology》1988,16(6):337-341
The effectiveness of a buffered sodium oleate solution was evaluated for detaching bacteria from ruminal digesta samples. A response surface derived from an octagonal design was used to determine the pH and concentration combination for maximum detachment of total and cellulolytic bacteria. The total number of bacteria detached increased up to 81% over control with treatment of a pH 8.8 and 1.5% sodium oleate solution. The recovery of cellulolytic bacteria was decreased to 35% of control with treatment of a pH 9.0 and 0.1% sodium oleate solution. Attempts to improve the recovery of viable bacteria exposed to sodium oleate solutions were unsuccessful. This response surface design identified an optimal pH and concentration that were consistent with existing information regarding detachment of total bacteria, and suggested that sodium oleate, at the concentrations tested, was toxic to the cellulolytic population of the rumen. 相似文献
46.
David H. Sachs Chief Sharon Germana Mona El-Gamil Kenth Gustafsson Francois Hirsch Karen Pratt 《Immunogenetics》1988,28(1):22-29
Class II genes of miniature swine have been characterized by restriction fragment length polymorphism (RFLP) analysis and by analysis of a series of clones isolated from a lymphocyte genomic library. For RFLP analysis, DNA samples from three independent major histocompatibility complex homozygous lines and three intra-MHC recombinant lines were digested with a variety of restriction enzymes and analyzed in Southern blots using human cDNA probes for DP, DQ, DR, and DZ alpha genes, and DP, DQ, DR, and DO beta genes. One, or at most two, unique fragments were detected by hybridization with each of the human probes tested. In contrast, multiple bands (five to six for most enzymes examined) were detected by each of the human probes tested, the majority of which were found to cross-react with at least three of these probes under conditions of moderate stringency. Genomic DNA from the SLA
c
haplotype was cloned into an EMBL-3 bacteriophage vector, and the corresponding genomic library was screened with each of these human cDNA probes. The class II genes thereby isolated from this library showed characteristics consistent with those anticipated from the RFLP analysis. Thus, unique genes were obtained which showed no evidence of cross-hybridization, while genes showed extensive cross-hybridization and were frequently detected in the library by more than one human gene probe. These data are consistent with early evolutionary divergence of a genes, prior to mammalian speciation, and with continuing evolution of genes, with possible shared usage of these genes by different a loci. The data also imply that genes can readily be assigned to loci homologous to their human counterparts, but that genes will require further mapping and/or sequence analysis to confirm assignments. 相似文献
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48.
Summary Cytological changes associated with the final maturation, and dehiscence of the 16-grain compound pollen (polyads) have been followed in anthers at female and male phase of flowering. InAcacia retinodes, the transition from female to male phase takes approximately 24 h. The spherical generative cell at female phase is connected with the vegetative cell plasma membrane by a junction zone. This is sited adjacent to a germinal aperture, comprising wall ingrowths and membrane labyrinths. By male phase, the generative cell has elongated into a spindle-shape, and its surface is characteristically scalloped. The membrane labyrinths, especially those at the apertures, now contain masses of coated vesicles, implicated in the transport and secretion of proteins. Two-dimensional projections indicate that the generative cell and vegetative nucleus are closely associated forming a male germ unit. 相似文献
49.
Development and characterization of continuous avian cell lines depleted of mitochondrial DNA 总被引:6,自引:0,他引:6
Réjean Morais Paul Desjardins Chanta Turmel Karen Zinkewich-Péotti 《In vitro cellular & developmental biology. Plant》1988,24(7):649-658
Summary Populations of quail and chicken cells were treated with ethidium bromide, an inhibitor of mitochondrial DNA replication.
After long-term exposure to the drug, the cell populations were transferred to ethidium bromide (EtdBr)-free medium, and cloned.
Clones HCF7 (quail) and DUS-3 (chicken) were propagated for more than a year, and then characterized. Analysis of total cellular
DNA extracted from these cells revealed no characteristic mitochondrial DNA molecule by Southern blot hybridization of HindIII-
or AvaI-digested total cellular DNA probed with cloned mitochondrial DNA fragments. Reconstruction experiments, where a small
number of parental cells was mixed with HCF7 cells and DUS-3 cells before extraction of total cellular DNA, further strengthen
the notion that the drug-treated cells are devoid of mitochondrial DNA molecules. The cell populations were found to proliferate
at a moderately reduced growth rate as compared to their respective parents, to be auxotrophic for uridine, and to be stably
resistant to the growth inhibitory effect of EtdBr and chloramphenicol. At the ultrastructural level, mitochondria were considerably
enlarged and there was a severe reduction in the number of cristae within the organelles and loss of cristae orientation.
Morphometric analysis revealed a fourfold increase of the mitochondrial profile area along with a twofold decrease of the
numerical mitochondrial profiles. Analysis of biochemical parameters indicated that the cells grew with mitochondria devoid
of a functional respiratory chain. The activity of the mitochondrial enzyme dihydroorotate dehydrogenase was decreased by
95% and presumably accounted for uridine auxotrophy.
This work was supported by a grant from the Medical Research Council of Canada. 相似文献
50.