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71.
胸腺细胞经ProTα和/或氢化考的松处理以后,采用PI染色法检测亚二倍体细胞百分率、荧光光度计检测细胞内游离Ca~(2 )浓度及琼脂糖凝胶电泳定性检验DNA片段化。结果发现单独或与氢化考的松联合应用,ProTα均可明显促进DNA片断化、提高亚二倍体细胞百分率并且也明显提高细胞内游离Ca~(2 )浓度。本实验说明ProTα能促进胸腺细胞的凋亡。 相似文献
72.
鸡减蛋综合征病毒(EDSV—76)末端前体蛋白的基因结构分析 总被引:1,自引:0,他引:1
从中国发病鸡群中分离的鸡减蛋综合征病毒弱毒株AA-2,经常规方法提取其病毒核酸后,组建了完整的限制性内切酶PstI及HingⅢ水解片段的基因文库,并对其中HindⅢ,-SacⅠ进行了序列测定。同源比较分析证明:其L链含编码病毒末端前体蛋白,容量为580个氨基酸残基的开放读码框架。 相似文献
73.
球孢白僵菌高渗适应性相关基因Bbmpd的克隆与表达分析 总被引:1,自引:0,他引:1
【目的】克隆与球孢白僵菌(Beauveria bassiana)的高渗适应性相关基因,并对其功能进行分析,以揭示球孢白僵菌对高渗等逆境适应的分子机理。【方法】利用YADE法克隆T-DNA的侧翼序列并进行基因组步行,获得突变基因的全长及上游序列;利用RT-PCR技术分析突变基因的表达特性以及与Bbhog1的关系;采用同源重组技术敲除Bbmpd基因。【结果】克隆得到插入突变基因及其上、下游序列全长3037bp。该基因与编码球孢白僵菌的1-磷酸甘露醇脱氢酶基因相似性为98%。Bbmpd的表达受高渗环境(0.8mol/L NaCl)的诱导,受Bbhog1信号途径的激活调节,Bbhog1缺失导致Bbmpd表达下调。Bbmpd缺失突变体在高渗胁迫下的生长受到明显抑制。Bbmpd缺失不影响球孢白僵菌在查氏培养基上的生长和产孢。【结论】由T-DNA突变体克隆了编码球孢白僵菌1-磷酸甘露醇脱氢酶基因Bbmpd,该基因的表达受高渗环境的诱导和Bbhog1的调控,与球孢白僵菌高渗适应性相关。 相似文献
74.
应用噬菌体抗体库技术制备全人源抗滋养层细胞表面抗原-2(Trop-2)特异性Fab抗体片段.抗体库经细胞筛选和固相抗原筛选,获得特异性的阳性克隆.阳性载体经核酸序列分析后,构建工程菌,经IPTG诱导表达,SDS-PAGE和Western blot分析,呈现28 ku和32 ku大小的两条蛋白质条带.Fab分子经流式细胞术、细胞免疫荧光检测,结果表明,Fab能够与BxPc3细胞膜蛋白特异性结合,而与NIH3T3细胞不结合.免疫共沉淀与质谱分析结果表明,该Fab分子能够与Trop-2蛋白特异性结合.免疫组化显示,该抗体可结合胰腺癌细胞膜蛋白,在细胞培养液中加入Fab,能够抑制BxPc3细胞的生长.以上研究结果提示,该抗体有望成为胰腺癌临床影像诊断或治疗的候选分子. 相似文献
75.
Yoon-Jeong Park Jin Chang Pen-Chung Chen Victor Chi-Min Yang 《Biotechnology and Bioprocess Engineering》2001,6(5):326-331
With the aim of developing a pH-sensitive controlled drug release system, a poly (L-lysine) (PLL) based cationic semi-interpenetrating polymer network (semi-IPN) has been synthesized. This cationic hydrogel
was designed to swell at lower pH and de-swell at higher pH and therefore be applicable for achieving regulated drug release
at a specific pH range. In addition to the pH sensitivity, this hydrogel was anticipated to interact with an ionic drug, providing
another means to regulate the release rate of ionic drugs. This semi-IPN hydrogel was prepared using a free-radical polymerization
method and by crosslinking of the polyethylene glycol (PEG)-methacrylate polymer through the PLL network. The two polymers
were penetrated with each other via interpolymer complexation to yield the semi-IPN structures. The PLL hydrogel thus prepared
showed dynamic swelling/de-swelling behavior in response to pH change, and such a behavior was influenced by both the concentrations
of PLL and PEG-methacrylate. Drug release from this semi-IPN hydrogel was also investigated using a model protein drug, streptokinase.
Streptokinase release was found to be dependent on its ionic interaction with the PLL backbones as well as on the swelling
of the semi-IPN hydrogel. These results suggest that a PLL semi-IPN hydrogel could potentially be used as a drug delivery
platform to modulate drug release by pH-sensitivity and ionic interaction. 相似文献
76.
77.
An Appropriate Cutoff Value for Determining the Colonization of Helicobacter pylori by the Pyrosequencing Method: Comparison with Conventional Methods 下载免费PDF全文
78.
Se Ryun Kwon Yue Jai Kang Dong Jin Lee Eun Hye Lee Yoon Kwon Nam Sung Koo Kim Ki Hong Kim 《Molecular biotechnology》2009,42(2):154-159
Vibrio anguillarum ghosts (VAG) were generated, for the first time, using a conjugation vector containing a ghost bacteria inducing cassette,
pRK-λPR-cI-Elysis, in which the expression of PhiX174 lysis gene E was controlled by the P
R
/cI regulatory system of lambda phage. By scanning electron microscopy, holes ranging 80–200 nm in diameter were observed in
the VAG. To avoid the presence of bacterial genomic DNA and an antibiotic resistance gene in the final VAG product, we constructed
a new dual vector, pRK-λPR-cI-E-SNA, containing the E-mediated lysis cassette and the staphylococcal nuclease A (SNA)-mediated DNA degradation cassette, and generated safety-enhanced VAG for use as a fish vaccine. 相似文献
79.
80.
Jin SM Lazarou M Wang C Kane LA Narendra DP Youle RJ 《The Journal of cell biology》2010,191(5):933-942
PINK1 is a mitochondrial kinase mutated in some familial cases of Parkinson's disease. It has been found to work in the same pathway as the E3 ligase Parkin in the maintenance of flight muscles and dopaminergic neurons in Drosophila melanogaster and to recruit cytosolic Parkin to mitochondria to mediate mitophagy in mammalian cells. Although PINK1 has a predicted mitochondrial import sequence, its cellular and submitochondrial localization remains unclear in part because it is rapidly degraded. In this study, we report that the mitochondrial inner membrane rhomboid protease presenilin-associated rhomboid-like protein (PARL) mediates cleavage of PINK1 dependent on mitochondrial membrane potential. In the absence of PARL, the constitutive degradation of PINK1 is inhibited, stabilizing a 60-kD form inside mitochondria. When mitochondrial membrane potential is dissipated, PINK1 accumulates as a 63-kD full-length form on the outer mitochondrial membrane, where it can recruit Parkin to impaired mitochondria. Thus, differential localization to the inner and outer mitochondrial membranes appears to regulate PINK1 stability and function. 相似文献