全文获取类型
收费全文 | 112920篇 |
免费 | 8821篇 |
国内免费 | 9775篇 |
专业分类
131516篇 |
出版年
2024年 | 259篇 |
2023年 | 1488篇 |
2022年 | 3437篇 |
2021年 | 5750篇 |
2020年 | 3969篇 |
2019年 | 4928篇 |
2018年 | 4660篇 |
2017年 | 3414篇 |
2016年 | 4789篇 |
2015年 | 6938篇 |
2014年 | 8204篇 |
2013年 | 8686篇 |
2012年 | 10421篇 |
2011年 | 9380篇 |
2010年 | 5800篇 |
2009年 | 5234篇 |
2008年 | 5983篇 |
2007年 | 5397篇 |
2006年 | 4688篇 |
2005年 | 3690篇 |
2004年 | 3167篇 |
2003年 | 2919篇 |
2002年 | 2427篇 |
2001年 | 2004篇 |
2000年 | 1788篇 |
1999年 | 1756篇 |
1998年 | 1093篇 |
1997年 | 1044篇 |
1996年 | 982篇 |
1995年 | 856篇 |
1994年 | 818篇 |
1993年 | 638篇 |
1992年 | 835篇 |
1991年 | 637篇 |
1990年 | 478篇 |
1989年 | 456篇 |
1988年 | 361篇 |
1987年 | 357篇 |
1986年 | 270篇 |
1985年 | 287篇 |
1984年 | 159篇 |
1983年 | 164篇 |
1982年 | 102篇 |
1981年 | 86篇 |
1980年 | 60篇 |
1979年 | 77篇 |
1977年 | 59篇 |
1975年 | 56篇 |
1974年 | 52篇 |
1973年 | 56篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
991.
Discerning an effective balance between equine infectious anemia virus attenuation and vaccine efficacy 总被引:2,自引:0,他引:2 下载免费PDF全文
Craigo JK Li F Steckbeck JD Durkin S Howe L Cook SJ Issel C Montelaro RC 《Journal of virology》2005,79(5):2666-2677
Among the diverse experimental vaccines evaluated in various animal lentivirus models, live attenuated vaccines have proven to be the most effective, thus providing an important model for examining critical immune correlates of protective vaccine immunity. We previously reported that an experimental live attenuated vaccine for equine infectious anemia virus (EIAV), based on mutation of the viral S2 accessory gene, elicited protection from detectable infection by virulent virus challenge (F. Li et al., J. Virol. 77:7244-7253, 2003). To better understand the critical components of EIAV vaccine efficacy, we examine here the relationship between the extent of virus attenuation, the maturation of host immune responses, and vaccine efficacy in a comparative study of three related attenuated EIAV proviral vaccine strains: the previously described EIAV(UK)DeltaS2 derived from a virulent proviral clone, EIAV(UK)DeltaS2/DU containing a second gene mutation in the virulent proviral clone, and EIAV(PR)DeltaS2 derived from a reference avirulent proviral clone. Inoculations of parallel groups of eight horses resulted in relatively low levels of viral replication (average of 10(2) to 10(3) RNA copies/ml) and a similar maturation of EIAV envelope-specific antibody responses as determined in quantitative and qualitative serological assays. However, experimental challenge of the experimentally immunized horses by our standard virulent EIAV(PV) strain by using a low-dose multiple exposure protocol (three inoculations with 10 median horse infective doses, administered intravenously) revealed a marked difference in the protective efficacy of the various attenuated proviral vaccine strains that was evidently associated with the extent of vaccine virus attenuation, time of viral challenge, and the apparent maturation of virus-specific immunity. 相似文献
992.
Human saliva contains a large number of proteins and peptides (salivary proteome) that help maintain homeostasis in the oral cavity. Global analysis of human salivary proteome is important for understanding oral health and disease pathogenesis. In this study, large-scale identification of salivary proteins was demonstrated by using shotgun proteomics and two-dimensinal gel electrophoresis-mass spectrometry (2-DE-MS). For the shotgun approach, whole saliva proteins were prefractionated according to molecular weight. The smallest fraction, presumably containing salivary peptides, was directly separated by capillary liquid chromatography (LC). However, the large protein fractions were digested into peptides for subsequent LC separation. Separated peptides were analyzed by on-line electrospray tandem mass spectrometry (MS/MS) using a quadrupole-time of flight mass spectrometer, and the obtained spectra were automatically processed to search human protein sequence database for protein identification. Additionally, 2-DE was used to map out the proteins in whole saliva. Protein spots 105 in number were excised and in-gel digested; and the resulting peptide fragments were measured by matrix-assisted laser desorption/ionization-mass spectrometry and sequenced by LC-MS/MS for protein identification. In total, we cataloged 309 proteins from human whole saliva by using these two proteomic approaches. 相似文献
993.
大型绿藻浒苔转化表达系统选择标记的筛选 总被引:1,自引:0,他引:1
主要研究了条浒苔对抗生素氯霉素和除草剂Basta的敏感性,以确定适合的阳性选择标记基因。应用不同浓度氯霉素(0、25、50、75、100、125μg/ml)和Basta(0、5、12.5、25、37.5、50μg/ml)对不同发育时期条浒苔细胞存活率影响进行了测定。实验结果表明:不同发育时期条浒苔对氯霉素和Basta的敏感性不同。其中最大浓度125μg/ml浓度的氯霉素在15d内对条浒苔孢子和小苗两个不同发育时期的细胞均难以达到全部杀死效果,相对存活率仍分别为1%和20%;而Basta对条浒苔孢子和小苗均具有很强的杀生作用,其中5μg/ml浓度的Basta在3d内可将条浒苔孢子全部杀死,12.5μg/ml浓度下约一周时间可以将浒苔小苗全部致死。本实验结果提示bar基因有可能成为浒苔基因工程较理想的选择标记基因。 相似文献
994.
995.
根据GenBank发布的绵羊GDF9基因外显子2的序列设计4对引物,采用PCR-SSCP技术分析GDF9基因外显子2在甘肃内羊新品种选育群羊中的单核苷酸多态性,并与产羔性状进行关联分析.结果表明,GDF9基因的扩增片段在所检测的新品种群羊中存在PCR-SSCP多态性,检测到3种基因型(AA、AB和BB),而在32只无角陶赛特母羊群中只检测到AA和AB基因型.测序结果显示,GDF9基因编码区第978位碱基发生A→G突变,但没有导致氨基酸的改变;第994位碱基发生G→A突变,导致Ⅴ变成Ⅰ(缬氨酸→异亮氨酸).新品种选育群羊产羔数的最小二乘均值关系为AB> AA> BB,统计分析结果初步表明3种基因型之间差异不显著(P>0.05).故该区域可能不是影响新品种群羊繁殖力的功能结构区城. 相似文献
996.
997.
998.
Y Liang Y Li Z Li Z Liu Z Zhang S Chang J Wu 《The international journal of biochemistry & cell biology》2012,44(11):1750-1760
Deficiencies in maternal diet, such as inadequate intake of folate, can inhibit normal development and lead to developmental defects. MicroRNAs (miRNAs) may play a role in mediating the effects of folate deficiency in the growing mammalian embryo, although conclusive evidences to support that possibility are not yet available. The goal of the present study was to investigate whether and how folate deprivation alters the properties of mouse embryonic stem cells (mESCs) in culture. For this purpose, mESCs were cultured in folate-deficient or complete culture medium. The results show that folate-deficient mESCs have a significantly higher rate of apoptosis, accumulate in G0/G1 and fail to proliferate. Expression profiling revealed several miRs and many mRNAs are differently expressed in folate-deficient cells. RT-PCR data confirmed differential expressions of 12 miRNAs in folate-deficient cells. Furthermore, bioinformatics analyses and in vitro studies suggested that miR-302a plays a critical role in mediating the effects of folate on cell proliferation and cell cycle-specific apoptosis by targeting Lats2 gene. Together, these results suggest that the effects of folate deficiency on mammalian development may be mediated by miRNAs that regulate proliferation and/or cell cycle progression in ESCs. 相似文献
999.
为研究植物病毒在RNA水平上对寄主植物的基因表达产生的影响,采用离子对高效液相色谱法,对黄瓜花叶病毒侵染烟草造成的总RNA的组成成分变化进行研究。离子对液相色谱法,是近几年才应用于对RNA进行分离、纯化和分析的一种试验技术,具有操作简单、重复性好、所需时间短等优点。选择适宜的离子对试剂,并对选定的离子对试剂正己胺/1,1,1,3,3,3-六氟-2-丙醇进行了优化,达到了较好的总RNA分离效果,并观察到健康植株和染病植株分离峰之间的差异,有差异的RNA种类还需进一步试验来验证。为研究植物病毒的致病机制提供一种新的试验方法和途径。 相似文献
1000.
Functional & Integrative Genomics - DNAJ proteins function as co-chaperones of HSP70 and play key roles in cell physiology to promote protein folding and degradation, especially under... 相似文献