全文获取类型
收费全文 | 6407篇 |
免费 | 291篇 |
国内免费 | 229篇 |
出版年
2023年 | 58篇 |
2022年 | 109篇 |
2021年 | 112篇 |
2020年 | 107篇 |
2019年 | 152篇 |
2018年 | 179篇 |
2017年 | 110篇 |
2016年 | 146篇 |
2015年 | 215篇 |
2014年 | 423篇 |
2013年 | 503篇 |
2012年 | 375篇 |
2011年 | 464篇 |
2010年 | 373篇 |
2009年 | 256篇 |
2008年 | 334篇 |
2007年 | 319篇 |
2006年 | 251篇 |
2005年 | 232篇 |
2004年 | 218篇 |
2003年 | 201篇 |
2002年 | 171篇 |
2001年 | 92篇 |
2000年 | 108篇 |
1999年 | 128篇 |
1998年 | 115篇 |
1997年 | 116篇 |
1996年 | 82篇 |
1995年 | 88篇 |
1994年 | 79篇 |
1993年 | 59篇 |
1992年 | 61篇 |
1991年 | 52篇 |
1990年 | 39篇 |
1989年 | 31篇 |
1988年 | 33篇 |
1987年 | 17篇 |
1986年 | 17篇 |
1985年 | 30篇 |
1984年 | 73篇 |
1983年 | 44篇 |
1982年 | 52篇 |
1981年 | 51篇 |
1980年 | 50篇 |
1979年 | 46篇 |
1978年 | 35篇 |
1977年 | 29篇 |
1976年 | 22篇 |
1975年 | 20篇 |
1974年 | 28篇 |
排序方式: 共有6927条查询结果,搜索用时 62 毫秒
71.
Hiromi Maekawa Tomoko Nakagawa Yoko Uno Kenji Kitamura Chikashi Shimoda 《Molecular & general genetics : MGG》1994,244(5):456-464
When the fission yeastSchizosaccharomyces pombe is starved for nitrogen, the cells are arrested in the G1 phase, enter the G0 phase and initiate sexual development. Theste13 mutant, however, fails to undergo a G1 arrest when starved for nitrogen and since this mutant phenotype is not suppressed by a mutation in adenylyl cyclase (cyr1), it would appear thatste13
+ either acts independently of the decrease in the cellular cAMP level induced by starvation for nitrogen, or functions downstream of this controlling event. We have used functional complementation to clone theste13
+ gene from anS. pombe genomic library and show that its disruption is not lethal, indicating that, while the gene is required for sexual development, it is not essential for cell growth. Nucleotide sequencing predicts thatste13
+ should encode a protein of 485 amino acids in which the consensus motifs of ATP-dependent RNA helicases of the DEAD box family are completely conserved. Point mutations introduced into these consensus motifs abolished theste13
+ functions. The predicted Ste13 protein is 72% identical to theDrosophila melanogaster Me31B protein over a stretch of 391 amino acids. ME31B is a developmentally regulated gene that is expressed preferentially in the female germline and may be required for oogenesis. Expression of ME31B cDNA inS. pombe suppresses theste13 mutation. These two evolutionarily conserved genes encoding putative RNA helicases may play a pivotal role in sexual development. 相似文献
72.
在豚鼠腹腔神经节细胞,γ-氨基丁酸(GABA)可诱发“去极化-超极化”的双相反应,去极化伴膜电阻减小,超极化伴膜电阻增大,这种反应在低钙高镁液中仍然存在。蝇蕈醇可模拟GABA双相反应,但在诱发相同强度的去极化反应时,蝇蕈醇产生的超极化弱于GABA。荷包牡丹碱(100μmol/L)和印防已毒素(100—300μmol/L)能可逆地抑制GABA诱发的双相反应。Baclofen对神经节细胞的膜电位无明显影响。结果提示,GABA双相反应的去极化相由GABAA/Cl-通道受体复合物介导,而超极化相很可能由不同于经典的GABAA受体,但其药理学行为又由与之十分相近的另一种GABA受体介导。 相似文献
73.
Production of mouse monoclonal antibodies using a continuous cell culture fermenter and protein G affinity chromatography 总被引:2,自引:0,他引:2
A. Zamboni I. Giuntini D. Gianesello F. Maddalena F. Rognoni D. Herbst 《Cytotechnology》1994,16(2):79-87
The production of anti--fetoprotein monoclonal antibodies for diagnostic use was carried out in a stirred tank fermenter equipped with a double membrane stirrer for bubble free aeration and continuous medium perfusion. A serum-free medium supplemented with 4 mM L-glutamine and 2.0 g/l glucose with a protein content of only 780 g/ml was used for the production process. The harvested antibodies were concentrated 50-fold using a tangential ultrafiltration system and were then purified in a one step purification process by protein G affinity chromatography. The purity of the final product (90%) was controlled by SDS-polyacrylamide gel electrophoresis, gel exclusion chromatography and isoelectric focussing. For further quality controls of the product the immunoglobulin subclass and the isoelectric point were determined and the specificity of the purified mAb was tested by RIA using125I labelled -fetoprotein.1.87 g of purified monoclonal antibodies were produced (90% purity) within 2 weeks. It was found that the use of this type of stirred tank fermenter combined with a one step purification process using protein G affinity chromatography represents a suitable method for the fast production of medium scale quantities (500 mg–5 g) of monoclonal antibodies for diagnostic use.Abbreviations AFP
-Fetoprotein
- BSA
bovine serum albumine
- FCS
Fetal calf serum
- HRP
horseradish peroxidase
- OPD
o-phenylenediamine dihydrochloride
- I.P.
isoelectric point
- IEF
isoelectric focussing
- PBS
Phosphate buffered saline 相似文献
74.
A. M. Bajo L. G. Guijarro M. G. Juarranz P. Valenzuela P. Martinez J. C. Prieto 《Bioscience reports》1993,13(2):69-77
Vasoactive intestinal peptide (VIP) has been shown to stimulate adenylyl cyclase activity in human endometrial membranes. The effect was dependent on the time and temperature of incubation as well as on the concentration of endometrial membrane proteins in the medium. In the presence of 1 M GTP, half-maximal stimulation of adenylyl cyclase activity was observed at 25.0±7.0 nM VIP, whereas the maximal activity (at 1 M VIP)corresponded to an increase of about 140% with respect to basal values (7.5±0.6 pmol cyclic AMP/min/mg of protein). However, the maximal stimulation of adenylyl cyclase activity was obtained with helodermin (1 M) that increased the activity by 170% over the basal. The relative potency of VIP-related peptides upon the adenylyl cyclase activity was: helodermin (ED50=1.8±1.4 nM)>VIP(ED50=25.0±7.0 nM)>PHI (ED50=725.0±127.2 nM). Secretin had a faint effect upon the adenylyl cyclase activity and glucagon was completely inefficient at this level. The presence of s and i subunits of G proteins in human endometrium was detected by immunoblot. Preliminary results showed the presence of two classes of125I-VIP receptors in human endometrial membranes with the following stoichoimetric parameters: high affinity receptor (Kd=2.0 nM, binding capacity 0.1 pmol VIP/mg protein) and low affinity receptor (Kd=0.43 M, binding capacity 13.1 pmol VIP/mg protein). The present results together with the known presence of VIP in human uterus and the actions of this neuropeptide in the adjacent myometrial tissue support the idea that VIP and related peptides may have a role in human endometrium. 相似文献
75.
76.
The chromosomal DNA of four strains of Gardnerella vaginaliswere digested with rare cutting restriction enzymes and analyzedby pulsed-field gel electrophoresis (PFGE). The four strainsstudied were two clinical isolates (GVP 004 & GVP 007) andtwo American Type Culture Collection strains (ATCC 14018 &ATCC 14019). The restriction enzyme SfiI generated two DNA fragmentsof about 0.6 Mb and 1.1 Mb in all four strains giving a G. vaginalisgenome size of about 1.7 Mb. A similar genome size was calculatedutilizing two more GC-rich sequence specific restriction endonucleases,NotI and AscI. When digested with AscI, the chromosomal DNAof all four strains gave rise to 11 to 12 DNA fragments rangingbetween 0.01 Mb to 0.43 Mb. DNA from the two clinical isolateswere digested by NotI (yielding 7 to 9 fragments), while theDNA from the two ATCC strains were resistant to NotI digestion.In contrast to the clinical isolates, DNA from the two ATCCstrains gave an identical profile for all restriction endonucleasestested. From double digestion experiments, the two SfiI sitescould be localized on two AscI fragments. From these PFGE studies,it is concluded that the G. vaginalis genome is a circular DNAthat ranges between 1.67 Mb and 1.72 Mb in size. 相似文献
77.
Arthur M. Brown 《The Journal of membrane biology》1993,131(2):93-104
Summary Ion channels are signaling molecules and by them-selves perform no work. In this regard they are un like the usual membrane
enzyme effectors for G proteins. The pathways of G protein receptor, G protein and ion channels are, therefore, purely infor
mational in function. Because a single G protein may have several ion channels as effectors, the effects should be coordinated
and this seems to be the case. Inhibition of Ca2+ current and stimulation of K+ currents would have a greater impact than either alone. Additional flexibility is provided by spontane ous noise in the complexes
of G protein receptor, G protein, and ion channel. By having a non-zero setpoint, the range of control is extended and the
responses become bi-directional. 相似文献
78.
79.
Noboru Sueoka 《Journal of molecular evolution》1993,37(2):137-153
Using a general form of the directional mutation theory, this paper analyzes the effect of mutations in mutator genes on the
G + C content of DNA, the frequency of substitution mutations, and evolutionary changes (cumulative mutations) under various
degrees of selective constraints. Directional mutation theory predicts that when the mutational bias between A/T and G/C nucleotide
pairs is equilibrated with the base composition of a neutral set of DNA nucleotides, the mutation frequency per gene will
be much lower than the frequency immediately after the mutator mutation takes place. This prediction explains the wide variation
of the DNA G + C content among unicellular organisms and possibly also the wide intragenomic heterogeneity of third codon
positions for the genes of multicellular eukaryotes. The present analyses lead to several predictions that are not consistent
with a number of the frequently held assumptions in the field of molecular evolution, including belief in a constant rate
of evolution, symmetric branching of phylogenetic trees, the generality of higher mutation frequency for neutral sets of nucleotides,
the notion that mutator mutations are generally deleterious because of their high mutation rates, and teleological explanations
of DNA base composition.
Presented at the NATO Advanced Research Workshop onGenome Organization and Evolution, Spetsai, Greece, 16–22 September 1992 相似文献
80.
Abstract: Study of transmembrane signaling via G proteins has focused to a large extent upon investigations of individual G protein-linked receptor-effector systems. Agonist-induced desensitization and down-regulation of β-adrenergic receptors, for example, have been studied extensively and adopted as a general model for G protein-linked receptor regulation. This review focuses not only on agonist regulation of adrenergic receptor gene expression, but also on how agonists regulate opposing adrenergic receptor-mediated pathways. This important feature of G protein-mediated pathways, i.e., cross-regulation and integration of information among several pathways, will be discussed in the context of what has been learned in the adrenergic receptor-coupled pathways. 相似文献