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141.
F J Livesey 《Briefings in Functional Genomics and Prot》2003,2(1):31-36
One of the critical limitations of current microarray technologies for use in expression analyses is the relatively large amount of input RNA required to generate labelled cDNA populations for array analysis. In situations where RNA is limiting, the options for expression profiling are to increase cDNA labelling and hybridisation efficiency, or to use an amplification strategy to generate enough RNA/cDNA for use with a standard labelling method. Sample amplification approaches must preserve the representation of the relative abundances of the different RNAs within the starting population and must also be highly reproducible. This review evaluates current signal and sample amplification technologies, including those that can be used to generate labelled cDNA populations for array analysis from as little as a single cell. 相似文献
142.
K. Kimura H. Kobayashi 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1996,680(1-2):213-219
In a potassium phosphate-poly(ethylene glycol) (PEG) system, RNA partitioning was accompanied by adsorption at the interface, depending on the molecular mass. Low-molecular-mass RNA showed the typical partition behavior of a soluble substance. Conversely, high-molecular-mass RNA was significantly adsorbed at the interface in the potassium phosphate-PEG1500 system. The adsorbed amount was proportional to the added amount, regardless of the phase volume ratio. The partitioning of high-molecular-mass RNA showed an amount-ratio-dependent distribution like that of a particle. 相似文献
143.
Rachel Baltz Jean-Luc Evrard Val/'erie Bourdon Andr/'e Steinmetz 《Sexual plant reproduction》1996,9(5):264-268
The protein PLIM-1 (formerly SF3) from sunflower is expressed exclusively in mature, free pollen. It contains two LIM domains associated with an acidic C-terminus comprising six copies of the pentapeptide motif (A,T,S) (E,D) TQN. We have expressed the pollen protein as well as some of its mutant forms inEscherichia coli and have used the bacterially produced proteins to study interactions with nucleic acids. Our studies show that the protein binds DNA and RNA in vitro to form large complexes, while mutant polypeptides containing either a single LIM domain or a destabilized first or second LIM domain do not. Although these data suggest that the biological function of PLIM-1 involves interactions with nucleic acids, its role in pollen development remains unclear. 相似文献
144.
反义RNA技术在植物基因工程领域中的应用 总被引:3,自引:0,他引:3
周跃钢 《生物化学与生物物理进展》1996,23(4):297-301
反义RNA技术在植物基因工程领域中的应用包括:a.番茄和其他水果的成熟控制;b.植物的抗病性;c.改变花卉的颜色;d.植物淀粉合成的控制;e.油料植物种子中脂肪酸合成的控制;f.杂交种子生产中雄性不育性的控制;g.其他. 相似文献
145.
栝楼核糖核酸酶(RNase TCS)对U碱基具有高度的专一性,在无脲、pH3.5、50℃时,它几乎都在-NP ↓ U-处裂解RNA.它与RNase T1,U2和有限的碱水解一起,可用于直接的酶法RNA序列分析. 相似文献
146.
核糖核酸(RNA)与稀土铽离子在pH5.0~6.5条件下能形成具有较强荧光的络合物,其激发峰在288nm处,发射峰为494nm及545nm处.RNA在0.1~10mg/L范围内与其荧光强度呈线性关系,其检出限为6.0×10 ̄(-8)mol/L.腺苷酸,尿苷酸,胞苷酸对RNA的干扰比较小,因此可在其存在下选择性地测定RNA. 相似文献
147.
Jacqueline S. Knight Francisco Madueño Simon A. Barnes John C. Gray 《Molecular biotechnology》1996,6(3):335-345
The levels of individual photosynthetic proteins can be independently decreased by theAgrobacterium-mediated transformation of plants with antisens RNA constructs. Protocols for the introduction of such constructs intoAgrobacterium, theAgrobacterium-mediated transformation of tobacco leaf disks, and the screening and analysis of the transgenic plants produced are described. 相似文献
148.
In vitro and in vivo action of antisense RNA 总被引:3,自引:0,他引:3
The transient or permanent expression of antisense RNA represents one option to apply antisense techniques in biotechnology
and medical research. Despite the increasing importance and use of antisense nucleic acids as well as their significant antisense-specific
phenotypic effects in vivo, there is an obvious lack of explanation for the mechanism of their action. By studying naturally
occurring antisense RNA and analyzing their mechanism of action we attempt to learn more about the design, the use, and the
critical parameters of artificial antisense RNA. Attempts to derive models from biochemical and structural studies for the
interactions between antisense RNAs and their targets will be discussed. 相似文献
149.
用σ ̄(70)或σ ̄(38)和核心酶(E)组成的大肠杆菌RNA聚合酶全酶(Eσ)对四种启动子进行了体外转录。结果表明:lacUV5,rplJ主要被Eσ ̄(70)识别,katE主要被Eσ ̄(38)识别,fic在低盐浓度时被Eσ ̄(70)识别,在高浓度盐时被Eσ ̄(38)识别;Eσ ̄(38)转录特异性启动子所需酶量大于Eσ ̄(70)转录特异性启动子的酶量;在含有分别对σ ̄(70)或σ ̄(38)亲和性大的混和启动子的体外转录中,启动子之间不存在干扰和竞争,转录水平与启动子浓度成正相关。 相似文献
150.
Enzymatic labeling of nucleic acids is a fundamental tool in molecular biology with virtually every aspect of nucleic acid
hybridization technique involving the use of labeled probes. Different methods for enzymatic labeling of DNA, RNA and oligonucleotide
probes are available today. In this review, we will describe both radioactive and nonradioactive labeling methods, yet the
choice of system for labeling the probe depends on the application under study. 相似文献