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61.
S-亚硝基化是一种重要的蛋白质翻译后修饰方式, 是指一氧化氮(NO)基团共价连接至靶蛋白特定半胱氨酸残基的自由巯基, 从而形成S-亚硝基硫醇(SNO)的过程。S-亚硝基化修饰广泛存在于各有机体中, 通过改变蛋白质生化活性、稳定性、亚细胞定位以及蛋白质-蛋白质相互作用等机制而调控不同的生物学过程或信号通路。在蛋白质S-亚硝基化检测分析方法中, 最为广泛使用的是生物素转化法(biotin switch assay), 其基本原理是首先封闭未被修饰的自由巯基, 进而将被修饰的SNO基团特异地还原为自由巯基并使用生物素将其特异标记。被生物素标记的半胱氨酸残基(即被修饰位点)可进一步通过蛋白质免疫印迹和/或质谱等方法进行检测分析。该文详细描述了植物蛋白质样品的体内和体外生物素转化法的实验流程, 并对实验过程中的注意事项进行了讨论。 相似文献
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Induction of the second exopolysaccharide (EPSb) in Rhizobium meliloti SU47 by low phosphate concentrations. 总被引:1,自引:0,他引:1 下载免费PDF全文
In previous work, Rhizobium meliloti SU47 produced its alternative exopolysaccharide (EPSb [also called EPS II]) only in strains that were genetically altered to activate EPSb synthesis. Here we report that EPSb synthesis is not entirely cryptic but occurred under conditions of limiting phosphate. This was shown in several different exo mutants that are blocked in the synthesis of the normal exopolysaccharide, succinoglycan. In addition, EPSb biosynthetic gene expression was markedly increased by limiting phosphate. An apparent regulatory mutant that does not express alkaline phosphatase activity was unable to produce EPSb under these conditions. A mucR mutant that was previously shown to produce EPSb instead of the normal exopolysaccharide, succinoglycan, was not sensitive to phosphate inhibition of EPSb synthesis. No evidence was found to indicate that exoX, which affects succinoglycan synthesis, had any influence on EPSb synthesis. In contrast to limiting phosphate, limiting nitrogen or sulfur did not stimulate EPSb synthesis as it does succinoglycan. 相似文献
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Functional and evolutionary relatedness of genes for exopolysaccharide synthesis in Rhizobium meliloti and Rhizobium sp. strain NGR234. 总被引:4,自引:4,他引:0 下载免费PDF全文
Rhizobium meliloti SU47 and Rhizobium sp. strain NGR234 produce distinct exopolysaccharides that have some similarities in structure. R. meliloti has a narrow host range, whereas Rhizobium strain NGR234 has a very broad host range. In cross-species complementation and hybridization experiments, we found that several of the genes required for the production of the two polysaccharides were functionally interchangeable and similar in evolutionary origin. NGR234 exoC and exoY corresponded to R. meliloti exoB and exoF, respectively. NGR234 exoD was found to be an operon that included genes equivalent to exoM, exoA, and exoL in R. meliloti. Complementation of R. meliloti exoP, -N, and -G by NGR234 R'3222 indicated that additional equivalent genes remain to be found on the R-prime. We were not able to complement NGR234 exoB with R. meliloti DNA. In addition to functional and evolutionary equivalence of individual genes, the general organization of the exo regions was similar between the two species. It is likely that the same ancestral genes were used in the evolution of both exopolysaccharide biosynthetic pathways and probably of pathways in other species as well. 相似文献
66.
Two genes that regulate exopolysaccharide production in Rhizobium meliloti. 总被引:8,自引:5,他引:3 下载免费PDF全文
We describe a new Rhizobium meliloti gene, exoX, that regulates the synthesis of the exopolysaccharide, succinoglycan, exoX resembled the psi gene of R. leguminosarum bv. phaseoli and the exoX gene of Rhizobium sp. strain NGR234 in its ability to inhibit exopolysaccharide synthesis when present in multiple copies, exoX did not appear to regulate the expression of exoP. The effect of exoX was counterbalanced by another R. meliloti gene, exoF. exoF is equivalent to Rhizobium sp. strain NGR234 exoY and resembles R. leguminosarum bv. phaseoli pss2 in its mutant phenotype and in portions of its deduced amino acid sequence. The effect of exoF on the succinoglycan-inhibiting activity of exoX depended on the relative copy numbers of the two genes. exoX-lacZ fusions manifested threefold-higher beta-galactosidase activities in exoF backgrounds than in the wild-type background. exoX mutants produced increased levels of succinoglycan. However, the exoF gene was required for succinoglycan synthesis even in an exoX mutant background. exoF did not affect the expression of exoP. Strains containing multicopy exoX formed non-nitrogen-fixing nodules on alfalfa that resembled nodules formed by exo mutants defective in succinoglycan synthesis. exoX mutants formed nitrogen-fixing nodules, indicating that, if the inhibition of succinoglycan synthesis within the nodule is necessary for nitrogen fixation, then exoX is not required for this inhibition. We present indirect evidence that succinoglycan synthesis within the nodule is not necessary for bacteroid function. 相似文献
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近年来,转基因技术与产品在中国的传播遭遇了一定障碍.科学界和相关政府部门往往从科学知识的普及角度克服这些障碍.但实际上,转基因传播障碍的背后有着更为隐蔽的反智的社会态度及对传统农业的迷恋心理,三个因素都与中国传统直观外推思维方式有关.转基因传播所面临的实际是构造自然观与有机自然观之间的冲突.对有机自然观的误读与当前弥漫的反智主义关系密切,转基因带有“高科技”的突出特征,恰恰容易成为反智主义攻击的对象.在有机自然观和“反智”传统的影响下,社会上对传统农业的迷恋有着相当的市场.在此情况下,转基因技术要克服传播障碍,必须重构环境,除了以经济动力继续推动传播工作外,也要让技术发展与人们普遍接受的自然观相结合. 相似文献
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本文利用三维笛卡儿空间座标对菲氏叶猴(Presbytis phayrei)和黑叶猴(P.francoisi)颅骨的形态结构作了比较研究,描绘了它们的侧视和俯视投影图形。结果表明,两种投影图形均显示黑叶猴的颅骨大于菲氏叶猴,且显著性增大的区域出现在眉点、囟门、枕骨大孔附近和下颌骨体的高度。但菲氏叶猴较黑叶猴具有更凸的面颅。据枕骨隆突至眉点的侧视投影弧长,两种叶猴的数学模式均为:Y=a+bX-cX~2(R=0.92)。从侧视投影看,两种叶猴自枕骨隆突至眉点主要表出大小的不同。若据俯视投影图,两种颅骨的差异主要出现在外耳道以前的部分。黑叶猴较菲氏叶猴具有十分显著性增高的下颌骨体以及相对应更发达的咬肌和咀嚼器官,这很可能与它取食更多的果类食物有关。 相似文献