全文获取类型
收费全文 | 890篇 |
免费 | 53篇 |
国内免费 | 34篇 |
专业分类
977篇 |
出版年
2023年 | 5篇 |
2022年 | 4篇 |
2021年 | 24篇 |
2020年 | 9篇 |
2019年 | 23篇 |
2018年 | 19篇 |
2017年 | 26篇 |
2016年 | 12篇 |
2015年 | 26篇 |
2014年 | 33篇 |
2013年 | 39篇 |
2012年 | 20篇 |
2011年 | 31篇 |
2010年 | 27篇 |
2009年 | 40篇 |
2008年 | 40篇 |
2007年 | 51篇 |
2006年 | 53篇 |
2005年 | 43篇 |
2004年 | 38篇 |
2003年 | 25篇 |
2002年 | 40篇 |
2001年 | 34篇 |
2000年 | 33篇 |
1999年 | 16篇 |
1998年 | 11篇 |
1997年 | 15篇 |
1996年 | 16篇 |
1995年 | 20篇 |
1994年 | 16篇 |
1993年 | 16篇 |
1992年 | 18篇 |
1991年 | 14篇 |
1990年 | 15篇 |
1989年 | 9篇 |
1988年 | 13篇 |
1987年 | 15篇 |
1986年 | 6篇 |
1985年 | 17篇 |
1984年 | 19篇 |
1983年 | 8篇 |
1982年 | 15篇 |
1981年 | 9篇 |
1980年 | 2篇 |
1979年 | 6篇 |
1978年 | 1篇 |
1976年 | 1篇 |
1975年 | 1篇 |
1972年 | 1篇 |
1971年 | 1篇 |
排序方式: 共有977条查询结果,搜索用时 15 毫秒
71.
Shrikant Mali 《Indian journal of human genetics》2013,19(1):3-8
The structure of DNA was unraveled by Watson and Crick in 1953, and two decades later Arber, Nathans and Smith discovered DNA restriction enzymes, which led to the rapid growth in the field of recombinant DNA technology. From expressing cloned genes in bacteria to expressing foreign DNA in transgenic animals, DNA is now slated to be used as a therapeutic agent to replace defective genes in patients suffering from genetic disorders or to kill tumor cells in cancer patients. Gene therapy provides modern medicine with new perspectives that were unthinkable two decades ago. Progress in molecular biology and especially, molecular medicine is now changing the basics of clinical medicine. A variety of viral and non-viral possibilities are available for basic and clinical research. This review summarizes the delivery routes and methods for gene transfer used in gene therapy. 相似文献
72.
M. C. Pinto D. H. Campbell-Lendrum A. L. Lozovei U. Teodoro † C. R. Davies 《Medical and veterinary entomology》2001,15(2):132-139
Responses of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) to carbon dioxide (CO2) and human odour were investigated by field experiments in Parana State, southern Brazil. Catches of two predominant species: Lu. intermedia (Antunes & Coutinho) and Lu. whitmani Lutz & Neiva, were compared between traps baited with a human adult or with CO2 emitted at the human-equivalent rate. When the baits were only 40 cm apart, no difference of attractiveness was detected. When baits were separated by 20 m, however, significantly fewer sandflies (44% Lu. intermedia, 46% Lu. whitmani) were trapped with CO2 compared with human bait. This is the first field evidence that anthropophilic sandflies are attracted by human kairomones in addition to CO2. For both species [Lutzomyia intermedia and Lu. whitmani] [corrected], the proportion of human attractiveness attributable to CO2 was significantly more [corrected] for males than females; for Lu. intermedia males human bait was no more attractive than CO2 alone. Gender differences in sandfly olfactory sensitivity are likely to be associated with behavioural differences on the host, where females feed on blood and males find mates. With traps 20 m apart, both Lutzomyia spp. showed roughly linear increased responses (log-log scale) to 0.08-0.55% CO2 equivalent to 0.5-4 humans. This would explain why host size is generally proportional to attractiveness, as observed for other species of phlebotomine sandflies. 相似文献
73.
Wolk CP Fan Q Zhou R Huang G Lechno-Yossef S Kuritz T Wojciuch E 《Archives of microbiology》2007,188(6):551-563
The clones generated in a sequencing project represent a resource for subsequent analysis of the organism whose genome has
been sequenced. We describe an interrelated group of cloning vectors that either integrate into the genome or replicate, and
that enhance the utility, for developmental and other studies, of the clones used to determine the genomic sequence of the
cyanobacterium, Anabaena sp. strain PCC 7120. One integrating vector is a mobilizable BAC vector that was used both to generate bridging clones and
to complement transposon mutations. Upon addition of a cassette that permits mobilization and selection, pUC-based sequencing
clones can also integrate into the genome and thereupon complement transposon mutations. The replicating vectors are based
on cyanobacterial plasmid pDU1, whose sequence we report, and on broad-host-range plasmid RSF1010. The RSF1010- and pDU1-based
vectors provide the opportunity to express different genes from either cell-type-specific or -generalist promoters, simultaneously
from different plasmids in the same cyanobacterial cells. We show that pDU1 ORF4 and its upstream region play an essential
role in the replication and copy number of pDU1, and that ORFs alr2887 and alr3546 (hetF
A
) of Anabaena sp. are required specifically for fixation of dinitrogen under oxic conditions. 相似文献
74.
Germ line transmission and expression of an RNAi cassette in mice generated by a lentiviral vector system 总被引:1,自引:0,他引:1
We have used a lentiviral delivery system (LentiLox3.7) to generate transgenic mice harbouring RNA interference (RNAi) against
the hepatocyte nuclear factor 4 gamma (HNF4γ). HNF4γ is a nuclear receptor with unknown function. Our analyses performed on
founder (F0) and first generation (F1) mice revealed mosaicism in F0 founders and a low efficiency of transgenesis (6%) in F1 mice. These data, together with the observation of multiple silenced transgenes, do not favour the use of LentiLox3.7 lentivirus
for transgenesis. Despite the low efficiency of transgenesis, we achieved a tissue-dependent knockdown of HNF4γ expression
in some mice.
Milen Kirilov and Minqiang Chai contributed equally to this work. 相似文献
75.
Rescue and production of vaccine and therapeutic adenovirus vectors expressing inhibitory transgenes
Gall JG Lizonova A EttyReddy D McVey D Zuber M Kovesdi I Aughtman B King CR Brough DE 《Molecular biotechnology》2007,35(3):263-273
Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the
inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing
the inhibiting transgene product. A gene regulation system based on the tet operon was used to allow the rescue and efficient growth of adenovectors that express transgenes to high levels. A key advantage
to this system is that repression of transgene expression is mediated by the packaging cell line, thus, expression of regulatory
products from the adenovector are not required. This provides a simple, broadly applicable system wherein transgene repression
is constitutive during vector rescue and growth and there is no effect on adenovector-mediated expression of gene products
in transduced cells. Several high-level expression vectors based on first- and second-generation adenovectors were rescued
and produced to high titer that otherwise could not be grown. Yields of adenovectors expressing inhibitory transgene products
were increased, and the overgrowth of cultures by adenovectors with nonfunctional expression cassettes was prevented. The
gene regulation system is a significant advancement for the development of adenovirus vectors for vaccine and other gene transfer
applications. 相似文献
76.
77.
Michaela Šimčíková Kristala L. J. Prather Duarte M. F. Prazeres 《Biotechnology & genetic engineering reviews》2015,31(1-2):82-107
Despite very good safety records, clinical trials using plasmid DNA failed due to low transfection efficiency and brief transgene expression. Although this failure is both due to poor plasmid design and to inefficient delivery methods, here we will focus on the former. The DNA elements like CpG motifs, selection markers, origins of replication, cryptic eukaryotic signals or nuclease-susceptible regions and inverted repeats showed detrimental effects on plasmids’ performance as biopharmaceuticals. On the other hand, careful selection of promoter, polyadenylation signal, codon optimization and/or insertion of introns or nuclear-targeting sequences for therapeutic protein expression can enhance the clinical efficacy. Minimal vectors, which are devoid of the bacterial backbone and consist exclusively of the eukaryotic expression cassette, demonstrate better performance in terms of expression levels, bioavailability, transfection rates and increased therapeutic effects. Although the results are promising, minimal vectors have not taken over the conventional plasmids in clinical trials due to challenging manufacturing issues. 相似文献
78.
79.
Brad S Coates 《BMC genomics》2015,16(1)
Background
The movement of mobile elements among species by horizontal transposon transfer (HTT) influences the evolution of genomes through the modification of structure and function. Helitrons are a relatively new lineage of DNA-based (class II) transposable elements (TEs) that propagate by rolling-circle replication, and are capable of acquiring host DNA. The rapid spread of Helitrons among animal lineages by HTT is facilitated by shuttling in viral particles or by unknown mechanisms mediated by close organism associations (e.g. between hosts and parasites).Results
A non-autonomous Helitron independently annotated as BmHel-2 from Bombyx mori and the MITE01 element from Ostrinia nubilalis was predicted in the genomes of 24 species in the insect Order Lepidoptera. Integrated Helitrons retained ≥ 65% sequence identity over a 250 bp consensus, and were predicted to retain secondary structures inclusive of a 3′-hairpin and a 5′-subterminal inverted repeat. Highly similar Hel-2 copies were predicted in the genomes of insects and associated viruses, which along with a previous documented case of real-time virus-insect cell line transposition suggests that this Helitron has likely propagated by HTT.Conclusions
These findings provide evidence that insect virus may mediate the HTT of Helitron-like TEs. This movement may facilitate the shuttling of DNA elements among insect genomes. Further sampling is required to determine the putative role of HTT in insect genome evolution.Electronic supplementary material
The online version of this article (doi:10.1186/s12864-015-1318-6) contains supplementary material, which is available to authorized users. 相似文献80.
Gehl C Kaufholdt D Hamisch D Bikker R Kudla J Mendel RR Hänsch R 《The Plant journal : for cell and molecular biology》2011,67(3):542-553
Dynamic protein-protein interactions are essential in all cellular and developmental processes. Protein-fragment complementation assays allow such protein-protein interactions to be investigated in vivo. In contrast to other protein-fragment complementation assays, the split-luciferase (split-LUC) complementation approach facilitates dynamic and quantitative in vivo analysis of protein interactions, as the restoration of luciferase activity upon protein-protein interaction of investigated proteins is reversible. Here, we describe the development of a floated-leaf luciferase complementation imaging (FLuCI) assay that enables rapid and quantitative in vivo analyses of protein interactions in leaf discs floating on a luciferin infiltration solution after transient expression of split-LUC-labelled interacting proteins in Nicotiana benthamiana. We generated a set of eight Gateway-compatible split-LUC destination vectors, enabling fast, and almost fail-safe cloning of candidate proteins to the LUC termini in all possible constellations. We demonstrate their functionality by visualizing the well-established homodimerization of the 14-3-3 regulator proteins. Quantitative interaction analyses of the molybdenum co-factor biosynthesis proteins CNX6 and CNX7 show that the luciferase-based protein-fragment complementation assay allows direct real-time monitoring of absolute values of protein complex assembly. Furthermore, the split-LUC assay is established as valuable tool to investigate the dynamics of protein interactions by monitoring the disassembly of actin filaments in planta. The new Gateway-compatible split-LUC destination vector system, in combination with the FLuCI assay, provides a useful means to facilitate quantitative analyses of interactions between large numbers of proteins constituting interaction networks in plant cells. 相似文献