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61.
Specific Single or Double Proline Substitutions in the “Spring-loaded” Coiled-Coil Region of the Influenza Hemagglutinin Impair or Abolish Membrane Fusion Activity 下载免费PDF全文
Hui Qiao Sandra L. Pelletier Lucas Hoffman Jill Hacker R. Todd Armstrong Judith M. White 《The Journal of cell biology》1998,141(6):1335-1347
We tested the role of the “spring-loaded” conformational change in the fusion mechanism of the influenza hemagglutinin (HA) by assessing the effects of 10 point mutants in the region of high coiled-coil propensity, HA2 54–81. The mutants included proline substitutions at HA2 55, 71, and 80, as well as a double proline substitution at residues 55 and 71. Mutants were expressed in COS or 293T cells and assayed for cell surface expression and structural features as well as for their ability to change conformation and induce fusion at low pH. We found the following: Specific mutations affected the precise carbohydrate structure and folding of the HA trimer. All of the mutants, however, formed trimers that could be expressed at the cell surface in a form that could be proteolytically cleaved from the precursor, HA0, to the fusion-permissive form, HA1-S-S-HA2. All mutants reacted with an antibody against the major antigenic site and bound red blood cells. Seven out of ten mutants displayed a wild-type (wt) or moderately elevated pH dependence for the conformational change. V55P displayed a substantial reduction (~60– 80%) in the initial rate of lipid mixing. The other single mutants displayed efficient fusion with the same pH dependence as wt-HA. The double proline mutant V55P/ S71P displayed no fusion activity despite being well expressed at the cell surface as a proteolytically cleaved trimer that could bind red blood cells and change conformation at low pH. The impairment in fusion for both V55P and V55P/S71P was at the level of outer leaflet lipid mixing. We interpret our results in support of the hypothesis that the spring-loaded conformational change is required for fusion. An alternate model is discussed. 相似文献
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65.
“Song Lan” is a source of Chinese drugs such as “Daqingye”, “Banlangen”
or “Qingdai”. We have discovered that the two species, “Song Lan” (Isatis indigotica
Fort.) and woad (I. tinctoria L.), were mistakenly described in the literature due to their
morphological polymorphism. In order to clarify the two species, cytology examination, pollen analysis, electrophoretic analysis of isoenzymes and soluble protein were performed. The
results show that previous non-trichiferous type of woad is a “Song Lan”. As in woad,
“Song Lan” is also morphologically of great variability. The base of canline leaves in this
species may be sagittate or auriculate. We have not found the non-trichiferous type of woad
in our country. It is reported for the first time that the chromosome number for “Song
Lan” is 2n=14.
The content of the indole glucoside in fresh leaves of “Song Lan” is about five timeshigher than in woad. For medicine cultivation of “Song Lan” is favorable. 相似文献
66.
Geneticdivergenceanalysiswasconductedbasedonperformanceof12quantitativetraitsin85sunflowerinbredlines,tostudyitseffectivenessinpredictingyieldheterosisofthe72hybridsfromthem.Resultsshowedthatlinearregressionmodelfittherelationshipbetweenheterosisofseedoilcontentandgeneticdistance,andquadraticregressionequationtherelationshipbetweengrainyieldheterosisandgeneticdistance,whichcanbeusedinpredictingheterosisfromtheparentperformances.Clusteranalysiswaseffectivetocertainextent,butitsutilizationshouldbelimited.Grainyieldandoilcontentcanbeimprovedsimutaneouslyaccordingtotherelationsbetweengeneticdivergenceandtheheterosisofthesetwotraits. 相似文献
67.
小桐子愈伤组织的诱导 总被引:1,自引:0,他引:1
本文以小桐子的叶片、叶柄、茎段及下胚轴和子叶作为外植体,研究不同外植体类型对愈伤组织诱导的影响,结果表明叶柄的诱导率最高,其次为茎段的诱导率。同时以小桐子的下胚轴作为外植体,研究植物生长调节剂种类及浓度配比对愈伤组织诱导的影响,结果显示6-BA与2,4-D的组合更适宜小桐子愈伤组织的诱导,MS+6-BA0.5mg/L+2,4-D0.1mg/L为小桐子愈伤组织诱导的最佳培养基,其愈伤组织诱导率高达96.7%。本研究为小桐子愈伤组织的分化、植株再生及相关的遗传转化研究提供了参考。 相似文献
68.
目的:探讨Neuritin对大鼠脊髓损伤后神经元突起再生的作用.方法:分为Neuritin组,His组和假手术组.使用改良Allen's打击法打击Neuritin组和His组大鼠T10或T11节段脊髓.Neuritin组和His组经蛛网膜下腔置管局部连续给予Neuritin和His蛋白(6ug/d)一周.假手术组仅咬除椎板不损伤脊髓,经蛛网膜下腔置空管而部损伤脊髓.术后6h、3d、7d、14d、28d、56d分别观察:①运动功能评分(BBB评分)观察大鼠后肢运动功能恢复情况;②HE染色观察脊髓组织形态学变化;③免疫组织化学染色和Western blotting检测损伤段脊髓神经中丝蛋白(NF200)的修复与再生.结果:①BBB评分,Neuritin组和His组在一周内没有明显差别,但Neuritin组和His组的评分均低于假手术组,从术后第14d,实验组评分明显高于对照组(P<0.05);②HE染色可见损伤段脊髓出血、坏死及炎性细胞浸润;③免疫组化检测,Neuritin组的NF200平均光密度值(IOD/AREA)较His组明显增高(P<0.05);Western blotting检测的NF200灰度值(GMD)较His组明显增高(P<0.05),结论:持续外源性Neuritin能促进大鼠脊髓损伤后伤区神经元突起的再生,并能促进大鼠后肢运动功能的康复. 相似文献
69.
Xin-wei Wang Ai-sheng Xiong Quan-hong Yao Zhen Zhang Yu-shan Qiao 《Molecular biotechnology》2010,44(1):61-65
Low molecular weight RNA (LMW RNA) is generally obtained either from the total RNA or from total nucleic acids solution. Many
steps and chemical reagents are involved in traditional methods for LMW RNA isolation where degradation of LMW RNA often occurs,
especially for plant materials with high levels of secondary catabolites. In this study, an efficient method was developed
to directly isolate pure LMW RNA from pear peel, a material rich in polyphenolics that is covered with a layer of wax. The
method was based on polyethylene glycol (PEG) precipitation combining CTAB buffer which is often used to isolate RNA from
polysaccharide-rich and polyphenolics-rich materials. The entire procedure could be completed within 6 h and many samples
could be processed at the same time. Few and common chemicals are used with this method. Hence, it could be used as an ordinary
method in the laboratory. The developed method was further tested by isolating LMW RNA from Arabidopsis. Using the isolated LMW RNA samples, microRNAs were successfully detected and characterized. 相似文献
70.
Hai Jin He Xing Sheng Wang Rong Pan Liang Dong Wang Ming Nan Liu Rong Qiao He 《BMC cell biology》2009,10(1):81