全文获取类型
收费全文 | 26248篇 |
免费 | 2212篇 |
国内免费 | 3051篇 |
出版年
2024年 | 60篇 |
2023年 | 278篇 |
2022年 | 769篇 |
2021年 | 1327篇 |
2020年 | 903篇 |
2019年 | 1194篇 |
2018年 | 1146篇 |
2017年 | 856篇 |
2016年 | 1205篇 |
2015年 | 1659篇 |
2014年 | 2011篇 |
2013年 | 2144篇 |
2012年 | 2482篇 |
2011年 | 2248篇 |
2010年 | 1418篇 |
2009年 | 1288篇 |
2008年 | 1531篇 |
2007年 | 1328篇 |
2006年 | 1167篇 |
2005年 | 1038篇 |
2004年 | 866篇 |
2003年 | 840篇 |
2002年 | 637篇 |
2001年 | 457篇 |
2000年 | 401篇 |
1999年 | 428篇 |
1998年 | 266篇 |
1997年 | 236篇 |
1996年 | 203篇 |
1995年 | 170篇 |
1994年 | 164篇 |
1993年 | 123篇 |
1992年 | 134篇 |
1991年 | 131篇 |
1990年 | 72篇 |
1989年 | 77篇 |
1988年 | 51篇 |
1987年 | 41篇 |
1986年 | 32篇 |
1985年 | 42篇 |
1984年 | 30篇 |
1983年 | 22篇 |
1982年 | 11篇 |
1981年 | 5篇 |
1980年 | 3篇 |
1979年 | 5篇 |
1978年 | 3篇 |
1971年 | 2篇 |
1953年 | 1篇 |
1950年 | 1篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
71.
生物素可通过氨烷基磷酰胺基团连接到寡核苷酸5'末端,反应在水溶液中进行,核苷酸的侧链基团不用保护。我们以这种化学法合成了生物素标记的寡核苷酸探针,其显色灵敏度达50pg,杂交灵敏度达0.4fmol,并与酶标生物素寡核苷酸探针进行了比较,也对两种不同显色体系作了比较。 相似文献
72.
Separate functional domains of the herpes simplex virus type 1 protease: evidence for cleavage inside capsids. 总被引:5,自引:5,他引:0
下载免费PDF全文
![点击此处可从《Journal of virology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
B J Robertson P J McCann rd L Matusick-Kumar W W Newcomb J C Brown R J Colonno M Gao 《Journal of virology》1996,70(7):4317-4328
The herpes simplex virus type 1 (HSV-1) protease (Pra) and related proteins are involved in the assembly of viral capsids and virion maturation. Pra is a serine protease, and the active-site residue has been mapped to amino acid (aa) 129 (Ser). This 635-aa protease, encoded by the UL26 gene, is autoproteolytically processed at two sites, the release (R) site between amino acid residues 247 and 248 and the maturation (M) site between residues 610 and 611. When the protease cleaves itself at both sites, it releases Nb, the catalytic domain (N0), and the C-terminal 25 aa. ICP35, a substrate of the HSV-1 protease, is the product of the UL26.5 gene. As it is translated from a Met codon within the UL26 gene, ICP35 cd are identical to the C-terminal 329-aa sequence of the protease and are trans cleaved at an identical C-terminal site to generate ICP35 e,f and a 25-aa peptide. Only fully processed Pra (N0 and Nb) and ICP35 (ICP35 e,f) are present in B capsids, which are believed to be precursors of mature virions. Using an R-site mutant A247S virus, we have recently shown that this mutant protease retains enzymatic activity but fails to support viral growth, suggesting that the release of N0 is required for viral replication. Here we report that another mutant protease, with an amino acid substitution (Ser to Cys) at the active site, can complement the A247S mutant but not a protease deletion mutant. Cell lines expressing the active-site mutant protease were isolated and shown to complement the A247S mutant at the levels of capsid assembly, DNA packaging, and viral growth. Therefore, the complementation between the R-site mutant and the active-site mutant reconstituted wild-type Pra function. One feature of this intragenic complementation is that following sedimentation of infected-cell lysates on sucrose gradients, both N-terminally unprocessed and processed proteases were isolated from the fractions where normal B capsids sediment, suggesting that proteolytic processing occurs inside capsids. Our results demonstrate that the HSV-1 protease has distinct functional domains and some of these functions can complement in trans. 相似文献
73.
74.
Elumalai Sivamani Ping Shen Natacha Opalka Roger N. Beachy Claude M. Fauquet 《Plant cell reports》1996,15(5):322-327
The microprojectile bombardment of immature embryos has proven to be effective in transforming many indica rice varieties. One of the drawbacks of using immature embryos is the requirement of a large number of high quality immature embryos, which itself is a tedious and laborious process. To circumvent these problems, we have developed a procedure, using indica variety TN1 as a model that generates highly homogenous populations of embryogenic subcultured calli by selectively propagating a small number of regeneration-proficient calli derived from seeds. Thousands of embryogenic calli were produced from 50 seeds within 10 weeks. Ten to 20 independent R0 transgenic lines were regenerated per 500 embryogenic calli bombarded. The convenience and reliability offered by this transformation system has made transformation of indica rice a routine procedure.Abbreviations 2,4-D
2,4-dichlorophenoxy acetic acid
- NAA
naphthalene acetic acid
- BAP
6-benzylaminopurine
- kb
kilobase
- GUS
-glucuronidase
- X-gluc
5-bromo-4-chloro-3-indolyl--D-glucuronide
- HPH
hygromycin B phosphotransferase 相似文献
75.
大豆子叶内酸性磷酸酶活性的超微结构定位 总被引:6,自引:0,他引:6
开花后35~50 d 期间和萌发早期(播种后4~8 d)的大豆(Glycinem ax L.)种子中,酸性磷酸酶主要分布在子叶细胞中的蛋白体内;在内质网内也检测到酸性磷酸酶活性。此外,在萌发早期的部分子叶细胞的质膜外侧及其细胞壁基质中可见密集的酸性磷酸酶活性;而且在近质膜的胞质中常见到一些富含磷酸铅沉淀的胞质小泡,似与质膜融合 相似文献
76.
水稻幼芽细胞生物膜上的赤霉素结合蛋白的结合特性 总被引:1,自引:0,他引:1
在水稻 (Oryza sativa)幼芽中存在膜结合的赤霉素结合蛋白 ,其与 GA3 结合的平衡解离常数(Kd)为 6.5× 1 0 -8mol/ L,总浓度为 0 .3 pmol· mg-1 蛋白质。结合蛋白与 GA3 结合活力在 0℃时比 2 5℃时高 1 4 0 %。它与 GA3 结合的最适 p H为 5。 GA3 与此结合蛋白的结合量随反应时间延长而增加 ,1 h达最大值 ,以后又逐渐下降。 IAA、ABA可与 GA3 竞争赤霉素结合蛋白。 相似文献
77.
九种罕见的人类染色体异常核型报告 总被引:1,自引:1,他引:0
ReportonNineRareSpeciesofHumanChromosomalAbnormalKaryotypesHanWeitian;QuOu;YuPing;JiangMiao(LiaoningResearchInstituteofFamilyPlanning,Shenyang110031)自1983年以来,我们对数千例不育及胚胎丢失等生殖异常患者进行细胞遗传学研究,发现大量异常染色体核型,而且异常种类繁多,已报道世界首报人类异常核型25种[1]。最近,在不良妊娠患者中又发现9种异常核型,经湖南医科大学医学细胞遗传学国家培训中心鉴定,为世界首次报道.现报告如下。1病例摘要及核型例1男,30岁,表型及智力正常,其妻子妊娠两次,均在无任何诱因情… 相似文献
78.
79.
80.
Xianzhen Li Yunzhan Huang Degui Xu Dongping Xiao Fengxie Jin Peiji Gao 《Biotechnology letters》1996,18(2):205-210
Summary The cellobiose oxidizing enzyme of the newly isolated cellulolytic bacterium Cytophaga sp. LX-7 was produced extracellularly when grown on cellulose or other saccharides, which was previously noted only in fungi. The enzyme could use not only cellobiose, maltose, glucose and other saccharides but also cellulose as substrates, and use dichlorophenol indophenol and oxygen as electron acceptors. 相似文献