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981.
Requirement of Cysteines and Length of the Human Respiratory Syncytial Virus M2-1 Protein for Protein Function and Virus Viability 总被引:1,自引:0,他引:1
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The M2-1 protein of human respiratory syncytial virus (hRSV) promotes processive RNA synthesis and readthrough at RSV gene junctions. It contains four highly conserved cysteines, three of which are located in the Cys(3)-His(1) motif at the N terminus of M2-1. Each of the four cysteines, at positions 7, 15, 21, and 96, in the M2-1 protein of hRSV A2 strain was individually replaced by glycines. When tested in an RSV minigenome replicon system using beta-galactosidase as a reporter gene, C7G, C15G, and C21G located in the Cys(3)-His(1) motif showed a significant reduction in processive RNA synthesis compared to wild-type (wt) M2-1. C96G, which lies outside the Cys(3)-His(1) motif, was fully functional in supporting processive RNA synthesis in vitro. Each of these cysteine substitutions was introduced into an infectious antigenomic cDNA clone derived from hRSV A2 strain. Except for C96G, which resulted in a viable virus, no viruses were recovered with mutations in the Cys(3)-His(1) motif. This indicates that the Cys(3)-His(1) motif is critical for M2-1 function and for RSV replication. The functional requirement of the C terminus of the M2-1 protein was examined by engineering premature stop codons that caused truncations of 17, 46, or 67 amino acids from the C terminus. A deletion of 46 or 67 amino acids abolished the synthesis of full-length beta-galactosidase mRNA and did not result in the recovery of viable viruses. However, a deletion of 17 amino acids from the C terminus of M2-1 reduced processive RNA synthesis in vitro and was well tolerated by RSV. Relocation of the M2-1 termination codon upstream of the M2-2 initiation codons did not significantly affect the expression of the M2-2 protein. Both rA2-Tr17 and rA2-C96G did not replicate as efficiently as wt rA2 in HEp-2 cells and was restricted in replication in the respiratory tracts of cotton rats. 相似文献
982.
【目的】探究饮水中添加复合益生菌制剂(地衣芽孢杆菌、枯草芽孢杆菌和丁酸梭菌)对肉鸡肌肉品质的影响及作用机理。【方法】试验随机选取360只1日龄白羽肉仔鸡,随机分为3个处理组:对照组(CON),正常饮水;低剂量益生菌组(LG),饮水添加0.2%复合益生菌;高剂量益生菌组(HG),饮水添加0.4%复合益生菌,试验为期42d。【结果】与CON组相比,LG组和HG组显著增加肉鸡7、35和42 d平均体重,显著提高HG组肉鸡21-28、28-35、35-42阶段的平均日增重(P<0.05),LG组仅显著提高35-42阶段平均日增重;显著提高胸肌45 min、24 h、48 h红度和24 h黄度,降低24 h和48 h亮度及48 h滴水损失和蒸煮损失。HG组胸肌粗蛋白和粗脂肪含量显著高于CON组,而LG组差异不显著;两组均可降低胸肌灰分含量。添加复合益生菌可显著提高总抗氧化能力(total antioxidant capacity,T-AOC),而总超氧化物歧化酶(total-superoxide dismutase,T-SOD)、过氧化氢酶(catalase,CAT)含量有上升趋势;显著上... 相似文献
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985.
Behavioral responses to vibrational stimuli were examined in two subterranean termite species, Coptotermes formosanus Shiraki and Reticulitermes flavipes (Kollar). Termites habituated to vibrational stimulation when in social groups, but failed to do so individually, indicating
that habituation is a collective action. In assays on termite groups, both species demonstrated a similar pattern of behavioral
responses to vibrational stimuli: evanescent cessation of activity and movement, followed by withdrawal from the vibration
source. Groups of both species then gradually moved back toward the vibration source as a consequence of continuous exposure.
However, it took a significantly shorter period for C. formosanus to return (57 s) to the test arena and to resume (80 s) normal foraging activities in the test arena compared with R. flavipes, which took 97 and 227 s, respectively, when exposed to the vibration frequency of 120 bmpm. High vibration frequency (240
bmpm) increased the time required to return (C. formosanus, 80 s; R. flavipes, 153 s) and to resume regular locomotion (C. formosanus, 186 s; R. flavipes, 263 s). Our experiments demonstrate that workers play a crucial part in adjusting groups of termites to distressful vibrations.
Soldiers of R. flavipes demonstrated similar behavioral responses as workers, however, C. formosanus soldiers exhibited a transient positive response before withdrawal.
An erratum to this article is available at . 相似文献
986.
人松弛素原样蛋白H2在大肠杆菌中的表达、分离纯化及鉴定 总被引:4,自引:0,他引:4
利用聚合酶链式反应方法 ,从含有人前松弛素原cDNA基因的质粒 pMALp2X-hRLXH2上得到人松弛素原H2的编码基因 ,亚克隆到温度诱导型原核表达载体pBV2 2 0上 ,转化大肠杆菌DH5α细胞。经 4 2℃温度诱导 ,获得了目的基因的高效表达。目的蛋白质以包含体的形式存在于大肠杆菌细胞中。菌体经过超声波破碎 ,包含体裂解 ,蛋白质的体外变性还原 ,复性 ,SephadexG 75凝胶过滤层析 ,反相快速蛋白质液相层析等一系列分离纯化过程 ,得到了高纯度的重组Met 人松弛素原样蛋白H2 ,得率约为 2~ 3mg/L。对纯化的目的蛋白质进行了SDS PAGE电泳纯度分析 ,氨基酸组成分析 ,通过基质辅助激光解吸电离飞行时间质谱法测得目的蛋白质的分子量为 183 90 .4 (理论计算值为 183 92 .3 )。 相似文献
987.
988.
繁缕和无瓣繁缕六个居群的数值分析 总被引:6,自引:0,他引:6
对繁缕(Stellaria media)和无瓣繁缕(S.apetala)的6个居群的57个性状进行Q-聚类和R-聚类的研究。结果表明:(1)Q-聚类中,用一条结合线,可以把繁缕的4个居群聚为一类,无瓣繁缕的2个居群聚为一类。这一结果支持肥繁缕和无瓣繁缕划分为两个物种;(2)R-聚类中,发现了呈现完全正相关、极大正相关和极大负相关的性状,并根据R-聚类的结果,运用一条适当的结合线,把繁缕和无瓣繁缕的57个性状划为5个类群,并分析了各性状的分类学意义。 相似文献
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990.