全文获取类型
收费全文 | 50315篇 |
免费 | 2828篇 |
国内免费 | 5257篇 |
出版年
2023年 | 783篇 |
2022年 | 1082篇 |
2021年 | 1505篇 |
2020年 | 1430篇 |
2019年 | 1861篇 |
2018年 | 1483篇 |
2017年 | 1234篇 |
2016年 | 1277篇 |
2015年 | 1576篇 |
2014年 | 2483篇 |
2013年 | 3347篇 |
2012年 | 2174篇 |
2011年 | 2541篇 |
2010年 | 2020篇 |
2009年 | 2448篇 |
2008年 | 2637篇 |
2007年 | 2807篇 |
2006年 | 2716篇 |
2005年 | 2480篇 |
2004年 | 2274篇 |
2003年 | 2050篇 |
2002年 | 1844篇 |
2001年 | 1472篇 |
2000年 | 1199篇 |
1999年 | 1147篇 |
1998年 | 1046篇 |
1997年 | 896篇 |
1996年 | 898篇 |
1995年 | 825篇 |
1994年 | 776篇 |
1993年 | 575篇 |
1992年 | 531篇 |
1991年 | 442篇 |
1990年 | 388篇 |
1989年 | 296篇 |
1988年 | 313篇 |
1987年 | 296篇 |
1986年 | 242篇 |
1985年 | 308篇 |
1984年 | 389篇 |
1983年 | 281篇 |
1982年 | 316篇 |
1981年 | 240篇 |
1980年 | 259篇 |
1979年 | 241篇 |
1978年 | 196篇 |
1977年 | 149篇 |
1976年 | 141篇 |
1975年 | 114篇 |
1974年 | 117篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
161.
J. T. M. Koumans H. A. Akster G. J. Dulos J. W. M. Osse 《Cell and tissue research》1990,261(1):173-181
Summary We have developed a method for the dissociation and purification of myosatellite cells from white epaxial muscle of carp. The dissociated myosatellite cells were identified by their morphology, their ultrastructure, the formation of multinucleated myotubes containing myofibrils and the immunocytochemical demonstration of desmin. Desmin and 5-bromo-2-deoxyuridine (BrdU) were used to identify terminally differentiated and proliferating myosatellite cells, respectively. The in vitro behavior of myosatellite cells dissociated from carp of 5 cm standard length differed from that described for myosatellite cells of mammals and birds. No substantial proliferation of the myosatellite cells could be observed. Most cells were differentiated (desmin-positive, BrdU-negative) 17 h after plating, regardless of the medium used. This indicates that the investigated white epaxial muscle of carp of 5 cm standard length contains subpopulations of myosatellite cells, arrested at various stages of differentiation. 相似文献
162.
The 6.5 kb HindIII DNA fragment of the Lactococcus lactis subsp. cremoris H2 plasmid pDI21 was cloned into Escherichia coli POP13 with NM1149, and also directly into Lactococcus lactis subsp. lactis 4125 using a newly-constructed broad host-range vector pFX1. Proteinase was experessed in both transformed organisms. The proteinase resembles a PI type since it preferentially degraded -casein. The restriction map of the 6.5 kb proteinase gene fragment has minor differences from those of published plamid proteinase genes. High-efficiency electroporation with pFX1 provides a direct approach for gene cloning in lactococci.Abbreviations cfu
colony forming units
- HEPES
N-[2-hydroxyethyl]piperazine-N-[2-ethanesulphonic acid]
Dedicated to Prof. Dr. G. Drews on the occasion of his 65th birthday 相似文献
163.
164.
抗阿特拉津转基因大豆植株后代的遗传分析 总被引:9,自引:0,他引:9
本试验用阿特拉津溶液涂抹、荧光诱导动力学检测、分子杂交等方法对抗阿特拉津转基因大豆植株的后代进行了鉴定,在第二代及第三代中检测到了抗性基因的存在,表明从龙葵中得到的此抗阿特拉津 psbA 基因不仅能导人大豆叶绿体基因组中获得表达,而且可以遗传到后代。 相似文献
165.
Noriaki Murakami Yoshiyuki Tanaka Kunio Takishima Yuzo Minobe Makoto Matsuoka Sei-Ichiro Kiyota Shin-ichi Hatanaka Katsuhiro Sakano 《Journal of plant research》1990,103(4):419-434
We isolated the small subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO SSu) from a fern,Asplenium cataractarum and determined its 34 N-terminal amino acid sequence. We obtained a cDNA clone that contains the entire coding region of
the SSu from the same fern species, using synthetic oligonucleotide probes derived from the above amino acid sequence. It
contains a 525 bp open reading frame capable of coding for a polypeptide with 174 amino acids, 31 bp 5′-and 206 bp 3′-noncoding
regions. It was also elucidated that the precursor to the SSu contains a transit peptide of 53 amino acid residues and a mature
protein of 121 residues. We compared the deduced amino acid sequence of the fern SSu with those of 11 other vascular plant
species (including gymnosperms, monocots and dicots). As low as 55% homology was observed between those of a fern and seed
plants. Constancy of the amino acid substitution rate in RuBisCO SSu was supported by our relative rate test. Amino acid substitution
rate per year per site for RuBisCO SSu was calculated to be 0.81×10−9 assuming that the separation between pteridophytes and seed plants arose 380 million years ago. 相似文献
166.
167.
168.
Summary
Escherichia coli Rl is an Ag+-resistant strain that, as we have shown recently, harbours at least two large plasmids, pJT1 (83 kb) and pJT2 (77 kb). Tn5-Mob was introduced into theE. coli Rl host replicon via conjugation on membrane filters. The transfer functions of plasmid RP4-4 were provided in this process and Tn5-Mob clones mated withE. coli C600 yielded Ag+-resistant transconjugants. This mobilization procedure allowed transfer and expression of pJT1 Ag+ resistance inE. coli C600. Prior to use of Tn5-Mob mobilization, it was not possible to transfer Ag+-resistant determinant(s) intoE. coli by conjugation or transformation including high-voltage electroporation.E. coli C600 containing PJTI and PJT2 displayed decreased accumulation of Ag+ similar toE. coli R1.E. coli C600 could not tolerate 0.1 and 0.5 mM Ag+, rapidly accumulated Ag+ and became non-viable. Tn5-Mob mobilization may be useful in the study of metal resistance in bacteria, especially in strains not studied for resistance mechanisms. 相似文献
169.
We have studied the mechanisms of breakdown of 2'-5' oligoadenylates. We monitored the time-courses of degradation of ppp(A2'p5')nA (dimer to tetramer) and of 5'OH-(A2'p5')nA (dimer to pentamer) in unfractionated L1210 cell extract. The 5' triphosphorylated 2'-5' oligoadenylates are converted by a phosphatase activity. However, 2'-5' oligoadenylates are degraded mainly by phosphodiesterase activity which splits the 2'-5' phosphodiester bond sequentially at the 2' end to yield 5' AMP and one-unit-shorter oligomers. The nonlinear least-squares curve-fitting program CONSAM was used to fit these kinetics and to determine the degradation rate constant of each oligomer. Trimers and tetramers, whether 5' triphosphorylated or not, are degraded at the same rate, whereas 5' triphosphorylated dimer is rapidly hydrolyzed and 5'-OH dimer is the most stable oligomer. The interaction between degradation enzymes and the substrate strongly depends on the presence of a 5' phosphate group in the vicinity of the phosphodiester bond to be hydrolyzed; indeed, when this 5' phosphate group is present, as in pp/pA2'p5'A/or A2'/p5'A2'p5'A/, affinity is high and maximal velocity is low. Such a degradation pattern can control the concentration of 2'-5' oligoadenylates active on RNAse L either by limiting their synthesis (5' triphosphorylated dimer is the primer necessary for the formation of longer oligomers) and/or by converting them into inhibitory (e.g., monophosphorylated trimer) or inactive (e.g., nonphosphorylated oligomers) molecules. 相似文献
170.
Purification and properties of theDrosophila zen protein 总被引:1,自引:0,他引:1
Summary The zen protein is encoded by the zerknullt gene required for normal early development inDrosophila. Like many regulatory proteins of this type, zen contains a 60 amino acid homeobox sequence. We have purified the zen protein
and studied its solution behavior and its interaction with DNA. The zen protein exists as a monomer in solution with a molecular
weight of about 40000. It binds specifically to a site about 900 bases upstream from thezen gene. Within this binding site DNase protection experiments indicate that binding is confined to two regions approximately
11 and 14 bases in length that are separated by about 30 base pairs. The protein concentration dependence of the binding curve
suggests that protein binding is non cooperative. 相似文献