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161.
Gene identification in a complex chromosomal continuum by local genomic cross-referencing 总被引:8,自引:0,他引:8
Zoya Avramova Alexander Tikhonov Phillip SanMiguel Young-Kwan Jin Changnong Liu Sung-Sick Woo Rod A. Wing Jeffrey L. Bennetzen 《The Plant journal : for cell and molecular biology》1996,10(6):1163-1168
Most higher plants have complex genomes containing large quantities of repetitive DNA interspersed with low-copy-number sequences. Many of these repetitive DNAs are mobile and have homology to RNAs in various cell types. This can make it difficult to identify the genes in a long chromosomal continuum. It was decided to use genic sequence conservation and grass genome co-linearity as tools for gene identification. A bacterial artificial chromosome (BAC) clone containing sorghum genomic DNA was selected using a maize Adh1 probe. The 165 kb sorghum BAC was tested for hybridization to a set of clones representing the contiguous 280 kb of DNA flanking maize Adh1. None of the repetitive maize DNAs hybridized, but most of the low-copy-number sequences did. A low-copy-number sequence that did cross-hybridize was found to be a gene, while one that did not was found to be a low-copy-number retrotransposon that was named Reina. Regions of cross-hybridization were co-linear between the two genomes, but closer together in the smaller sorghum genome. These results indicate that local genomic cross-referencing by hybridization of orthologous clones can be an efficient and rapid technique for gene identification and studies of genome organization. 相似文献
162.
Female mate choice and the benefits of this behavior are criticalaspects of Darwinian sexual selection, but they are seldom documentedbecause it is difficult to identify the male trait(s) that femalesmay be seeking. We conducted experiments with grasshoppers (Melanoplussangutnipes: Orthoptera, Acrididae) to examine this behavior.Males that feed more intensively and select a diet mix thatpermits greater food intake (food intake per body mass per time)in laboratory trials were preferentially selected by females.These better foraging males on average provide greater paternalinvestment (greater spermatophore mass) to the female, whichincreases her reproductive rate (eggs produced per body massper time). However, paternal investment may not entirely explainfemale choice of better foraging males, because these maleswere still selected even if they had their food intake restrictedor had been allowed to recently mate, which reduces spermatophoreproduction. Furthermore, males change their mating strategyin response to female choice and the foraging abilities of surroundingmales. Poorer foraging males attempt forcible copulation ratherthan displaying and allowing female choice. A male will facultativelyswitch between these strategies depending on the foraging abilitiesof the surrounding males. While females attempt to reject forciblecopulation, forcible copulation reduces the frequency with whichfemales successfully copulate with better foraging males. Therefore,males that are less "attractive" to females adopt alternativemating strategies to counter female choice which would excludethem from mating.[Behav Ecol 7: 438444 (1996)] 相似文献
163.
Herman Yeger Diane Forget Jennifer Alami Bryan R. G. Williams 《In vitro cellular & developmental biology. Animal》1996,32(8):496-504
Summary The temporal and spatial expression patterns of the Wilms tumor gene, WT1, were studied during the organogenesis of the mouse
kidneyin vitro. In situ hybridization and immunocytochemistry localized cellular expression of WT1 in whole kidney organ cultures to the induced
metanephric mesenchyme and developing podocytes. Organ cultures were further characterized immunocytochemically with antibodies
that specifically labeled the different tubular epithelial components and supporting mesenchyme of the developing nephrons.
In organ cultures, the WT1 expression pattern could be visualized in induced metanephric mesenchyme and entire cell cohorts
of differentiating podocytes. Expression of WT1 and cell specific markers were retained in short-term monolayer cultures of
dissociated kidneys. The development of the metanephric kidneyin vitro involves a highly restricted temporal and spatial cellular expression pattern of WT1 which closely follows that observed
in tissue sections from gestational kidney isolated during organogenesis in the mouse. 相似文献
164.
Wendy L. Picking Jennifer A. Mertz Mary E. Marquart William D. Picking 《Protein expression and purification》1996,8(4):401-408
Shigella flexneriand related enteropathogenic bacteria are important agents of bacillary dysentery, a potentially life-threatening illness for children in underdeveloped regions of the world. Onset of shigellosis stems fromS. flexneriinvasion of colonic epithelial cells, leading to localized cell death and inflammation. Invasion plasmid antigens (Ipa) B, C, and D are three secreted proteins encoded by the large virulence plasmid ofS. flexnerithat have been implicated as essential effectors of this cell invasion process. These proteins are expressed as part of theipaoperon and are among the major targets of the host immune response to shigellosis. Biochemical characterization of the Ipa invasins has been complicated by the fact they have not been purified in the quantities needed for detailedin vitroanalysis. Here we describe the first cloning, expression, and extensive purification of IpaB and IpaC fusion proteins fromEscherichia colifor use in dissecting of the protein biochemistry ofS. flexneripathogenesis. A variety of approaches were used to prepare significant quantities of these proteins in their soluble forms, including the use of different host cell lines, modification of bacterial growth conditions, and the use of alternative plasmid expression vectors. Now that these Ipa proteins are available in a highly pure form, it will be possible to initiate studies on their important biological and immunological properties as well as their recruitment into high-molecular-weight protein complexes. Together with IpaD (purified as part of a previous study), these purified proteins will be useful for: (a) exploring properties of the host immune response toS. flexneriinvasion, (b) elucidating the specific biochemical properties that lead to pathogen internalization, (c) analyzing the importance of specific Ipa protein complexes in host cell invasion, and (d) monitoring, or perhaps even augmenting, the efficacy of live oral vaccines in human trials. 相似文献
165.
The Best Gastric Site for Obtaining a Positive Rapid Urease Test 总被引:1,自引:0,他引:1
Jae Soon Woo Hala M. T. EI-Zimaity† Robert M. Genta † Mahmoud M. Yousfi David Y. Graham‡ 《Helicobacter》1996,1(4):256-259
Background Rapid urease tests (RUTs) provide a simple, sensitive method of detecting Helicobacter pylori infection.
Objectives. Our aim, therefore, was to determine whether the yield of detecting H. pylori infection by RUT varied depending on the site of gastric biopsy.
Materials and Methods. Gastric biopsies were obtained from 50 patients for RUT by use of hp fast (GI Supply, Camp Hill, PA). Biopsies were taken from the prepyloric greater curve antrum, from the gastric angle, and from the greater curve in mid-corpus. One biopsy specimen was placed in the RUT gel, and the biopsy from the adjacent mucosa was placed in formalin for subsequent histological evaluation by using the Genta stain. RUTs were examined and scored at intervals of 5, 10, 15, 30, and 45 minutes and after 1, 2, 4, and 24 hours.
Results. Fifty patients were entered in the test (150 RUTs), 32 having H. pylori infection. There were no false-positive RUTs (specificity, 100%). The gastric angle site was positive in 100%. The prepyloric site was positive in 87%, and the corpus site was positive in 84.4% ( p < .052 for angle or prepyloric antrum versus corpus). The most common pattern was for all to be positive (74%). The median time to positivity was similar with angle and prepyloric sites (37.5 and 60 minutes, respectively, p = not significant) and shorter than the corpus biopsy (180 minutes); ( p < .05 for angle or prepyloric antrum versus corpus).
Conclusion. The maximum probability for detecting H. pylori infection using a RUT is to obtain a biopsy from the gastric angle. To prevent missing a positive result when intestinal metaplasia is present, we recommend that (at a minimum) biopsies be taken from both the angle and the corpus. 相似文献
Objectives. Our aim, therefore, was to determine whether the yield of detecting H. pylori infection by RUT varied depending on the site of gastric biopsy.
Materials and Methods. Gastric biopsies were obtained from 50 patients for RUT by use of hp fast (GI Supply, Camp Hill, PA). Biopsies were taken from the prepyloric greater curve antrum, from the gastric angle, and from the greater curve in mid-corpus. One biopsy specimen was placed in the RUT gel, and the biopsy from the adjacent mucosa was placed in formalin for subsequent histological evaluation by using the Genta stain. RUTs were examined and scored at intervals of 5, 10, 15, 30, and 45 minutes and after 1, 2, 4, and 24 hours.
Results. Fifty patients were entered in the test (150 RUTs), 32 having H. pylori infection. There were no false-positive RUTs (specificity, 100%). The gastric angle site was positive in 100%. The prepyloric site was positive in 87%, and the corpus site was positive in 84.4% ( p < .052 for angle or prepyloric antrum versus corpus). The most common pattern was for all to be positive (74%). The median time to positivity was similar with angle and prepyloric sites (37.5 and 60 minutes, respectively, p = not significant) and shorter than the corpus biopsy (180 minutes); ( p < .05 for angle or prepyloric antrum versus corpus).
Conclusion. The maximum probability for detecting H. pylori infection using a RUT is to obtain a biopsy from the gastric angle. To prevent missing a positive result when intestinal metaplasia is present, we recommend that (at a minimum) biopsies be taken from both the angle and the corpus. 相似文献
166.
Richard Karban Gregory English-Loeb M. Andrew Walker Jennifer Thaler 《Experimental & applied acarology》1995,19(4):189-197
We observed the number of predatory mites (Phytoseiidae:Typhlodromus caudiglans) on the foliage of 20 North American species of grapes (Vitis spp) plus the domesticated EuropeanVitis vinifera, all grown in a common garden. We found relatively few phytophagous mites. The numbers of phytophagous mites were not correlated with the plant characteristics that we measured. We found approximately five times as many predatory mites as phytophagous mites and the numbers of these phytoseiid predators were not affected by the availability of prey. Similarly, numbers of phytoseiids were unaffected by plant gender and, hence, the availability of pollen, another source of food. The numbers of phytoseiids were not clustered according to the taxonomic grouping of the tested plant species. Leaf surface characteristics explained over 25% of the variance in the numbers of phytoseiids. Numbers of phytoseiids were positively associated with the density of vein hairs, the density of bristles in leaf axils, and the presence of leaf domatia. These results suggest that sheltered habitats rather than food availability may limit the numbers of phytoseiid mites on grapevines. 相似文献
167.
McMahon Jennifer M.; White Wanda L.B.; Sayre Richard T. 《Journal of experimental botany》1995,46(7):731-741
Cassava is the most agronomically important of the cyanogeniccrops. Linamarin, the predominant cyanogenic glycoside in cassava,can accumulate to concentrations as high as 500 mg kg1fresh weight in roots and to higher levels in leaves. Recently,the pathway of linamarin synthesis and the cellular site oflinamarin storage have been determined. In addition, the cyanogenicenzymes, linamarase and hydroxynitrile lyase, have been characterizedand their genes cloned. These results, as well as studies onthe organ- and tissue-specific localization of linamarase andhydroxy-nitrile lyase, allow us to propose models for the regulationof cyanogenesis in cassava. There remain, however, many unansweredquestions regarding the tissue-specific synthesis, transport,and accumulation of cyanogenic glycosides. The resolution ofthe sequestions will facilitate the development of food processing,biochemical and transgenic plant approaches to reducing thecyanogen content of cassava foods. Key words: Cyanide, cyanogenic glycosides, linamarin, cyanogens 相似文献
168.
Comparative chromosome painting between two marsupials: origins of an XX/XY1Y2 sex chromosome system
Roland Toder Rachel J. W. O’Neill Johannes Wienberg Patricia C. M. O’Brien Lucille Voullaire Jennifer A. Marshall-Graves 《Mammalian genome》1997,8(6):418-422
Cross-species chromosome painting was used to investigate genome rearrangements between tammar wallaby Macropus eugenii (2n = 16) and the swamp wallaby Wallabia bicolor (2n = 10♀/11♂), which diverged about 6 million years ago. The swamp wallaby has an XX female:XY1Y2 male sex chromosome system thought to have resulted from a fusion between an autosome and the small original X, not involving
the Y. Thus, the small Y1 should represent the original Y and the large Y2 the original autosome. DNA paints were prepared from flow-sorted and microdissected chromosomes from the tammar wallaby.
Painting swamp wallaby spreads with each tammar chromosome-specific probe gave extremely strong and clear signals in single-,
two-, and three-color FISH. These showed that two tammar wallaby autosomes are represented unchanged in the swamp wallaby,
two are represented by different centric fusions, and one by a tandem fusion to make the very long arms of swamp wallaby Chromosome
(Chr) 1. The large swamp wallaby X comprises the tammar X as its short arm, and a tandemly fused 7 and 2 as the long arm.
The acrocentric swamp wallaby Y2 is a 2/7 fusion, homologous with the long arm of the X. The small swamp wallaby Y1 is confirmed as the original Y by its painting with the tammar Y. However, the presence of sequences shared between the microdissected
tammar Xp and Y on the swamp wallaby Y2 implies that the formation of the compound sex chromosomes involved addition of autosome(s) to both the original X and Y.
We propose that this involved fusion with an ancient pseudoautosomal region followed by fission proximal to this shared region.
Received: 16 October 1996/Accepted: 30 January 1997 相似文献
169.
Xiangning Deng Jennifer Moran Neal G. Copeland Debra J. Gilbert Nancy A. Jenkins Paul Primakoff Patricia A. Martin-DeLeon 《Mammalian genome》1997,8(2):94-97
We have determined the chromosomal localization of the murine gene encoding the 68-kDa sperm adhesion molecule 1, Spam1 or Ph-20. Using two independent approaches, fluorescence in situ hybridization (FISH) and interspecific backcross analysis, we show
that Spam1 maps to proximal mouse Chromosome (Chr) 6. This map position is within the conserved linkage group corresponding to human
Chr 7q, where the human homolog, SPAM 1, has been shown to map previously. Genetic mapping shows the gene to be very closely
linked to Met, one of the most proximal loci on MMU 6. It thus places the gene near the centromere and the junction of the Rb(6.16)24Lub
and Rb(6.15)1Ald translocations. The essential role of the Spam1 sperm antigen in mouse sperm-egg interactions and its gene
location provide strong support for its candidacy as the gene involved in the dysfunction of mouse sperm bearing the Rb(6.16)24Lub
or Rb(6.15)1Ald translocation.
Received: 16 July 1996 / Accepted: 23 September 1996 相似文献
170.