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51.
Structure and function of the enhancer 3' to the human A gamma globin gene. 总被引:5,自引:0,他引:5 下载免费PDF全文
An enhancer is located immediately 3' to the A gamma globin gene. We have used DNase I footprinting to map the sites of interaction of nuclear proteins with the DNA sequences of this enhancer. Eight footprints were discovered, distributed over 600 base pairs of DNA. Three of these contain a consensus binding site for the erythroid specific factor GATA-I. Each of these GATA-1 sites had an enhancer activity when inserted into a reporter plasmid and tested in human erythroleukemia cells. Other footprints within the enhancer contained consensus binding sequences for the ubiquitous, positive regulatory proteins AP2 and CBP-1. An Sp1-like recognition sequence was also identified. Synthetic oligonucleotides encompassing two of the footprints generated a slowly migrating complex in gel mobility shift assays. The same complex forms on a fragment of the human gamma globin gene promoter extending from -260 to -200. The DNaseI footprint of this protein complex with the enhancer overlapped a sequence, AGGAGGA, found within the binding site for a protein that interacts with the chicken beta globin promoter and enhancer, termed the stage selector element. We propose that this complex of proteins may be involved in the human gamma globin promoter-enhancer interaction. 相似文献
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53.
Gallagher PG Sabatino DE Basseres DS Nilson DM Wong C Cline AP Garrett LJ Bodine DM 《The Journal of biological chemistry》2001,276(45):41683-41689
Ankyrin defects are the most common cause of hereditary spherocytosis (HS). In several kindreds with recessive, ankyrin-deficient HS, mutations have been identified in the ankyrin promoter that have been proposed to decrease ankyrin synthesis. We analyzed the effects of two mutations, -108T to C and -108T to C in cis with -153G to A, on ankyrin expression. No difference between wild type and mutant promoters was demonstrated in transfection or gel shift assays in vitro. Transgenic mice with a wild type ankyrin promoter linked to a human (A)gamma-globin gene expressed gamma-globin in 100% of erythrocytes in a copy number-dependent, position-independent manner. Transgenic mice with the mutant -108 promoter demonstrated variegated gamma-globin expression, but showed copy number-dependent and position-independent expression similar to wild type. Severe effects in ankyrin expression were seen in mice with the linked -108/-153 mutations. Three transgenic lines had undetectable levels of (A)gamma-globin mRNA, indicating position-dependent expression, and four lines expressed significantly lower levels of (A)gamma-globin mRNA than wild type. Two of four expressing lines showed variegated gamma-globin expression, and there was no correlation between transgene copy number and RNA level, indicating copy number-independent expression. These data are the first demonstration of functional defects caused by HS-related, ankyrin gene promoter mutations. 相似文献
54.
Evidence for independent recruitment of zeta-crystallin/quinone reductase (CRYZ) as a crystallin in camelids and hystricomorph rodents 总被引:3,自引:2,他引:1
Zeta-crystallin/quinone reductase (CRYZ) is an NADPH oxidoreductase
expressed at very high levels in the lenses of two groups of mammals:
camelids and some hystricomorph rodents. It is also expressed at very low
levels in all other species tested. Comparative analysis of the mechanisms
mediating the high expression of this enzyme/crystallin in the lens of the
Ilama (Lama guanacoe) and the guinea pig (Cavia porcellus) provided
evidence for independent recruitment of this enzyme as a lens crystallin in
both species and allowed us to elucidate for the first time the mechanism
of lens recruitment of an enzyme- crystallin. The data presented here show
that in both species such recruitment most likely occurred through the
generation of new lens promoters from nonfunctional intron sequences by the
accumulation of point mutations and/or small deletions and insertions.
These results further support the idea that recruitment of CRYZ resulted
from an adaptive process in which the high expression of CRYZ in the lens
provides some selective advantage rather than from a purely neutral
evolutionary process.
相似文献
55.
Sabatino DE Wong C Cline AP Pyle L Garrett LJ Gallagher PG Bodine DM 《The Journal of biological chemistry》2000,275(37):28549-28554
In red blood cells ankyrin (ANK-1) provides the primary linkage between the erythrocyte membrane skeleton and the plasma membrane. We have previously demonstrated that a 271-bp 5'-flanking region of the ANK-1 gene has promoter activity in erythroid, but not non-erythroid, cell lines. To determine whether the ankyrin promoter could direct erythroid-specific expression in vivo, we analyzed transgenic mice containing the ankyrin promoter fused to the human (A)gamma-globin gene. Sixteen of 17 lines expressed the transgene in erythroid cells indicating nearly position-independent expression. We also observed a significant correlation between the level of Ank/(A)gamma-globin mRNA and transgene copy number. The level of Ank/(A)gamma mRNA averaged 11% of mouse alpha-globin mRNA per gene copy at all developmental stages. The addition of the HS2 enhancer from the beta-globin locus control region to the Ank/(A)gamma-globin transgene resulted in Ank/(A)gamma-globin mRNA expression in embryonic and fetal erythroid cells in six of eight lines but resulted in absent or dramatically reduced levels of Ank/(A)gamma-globin mRNA in adult erythroid cells in eight of eight transgenic lines. These data indicate that the minimal ankyrin promoter contains all sequences necessary and sufficient for erythroid-specific, copy number-dependent, position-independent expression of the human (A)gamma-globin gene. 相似文献
56.
57.
Human lung fibroblasts (W138) can be brought to a quiescent state by removal of serum from the medium or by lowering of the extracellular Ca++. Upon return of Ca++ or serum, the cells enter the G1 phase and progress to S within 15–18 hours. Since multiple G1 phase blocks have been demonstrated, we wished to determine whether the Ca++ and serum block were equivalent since previous data suggested that these two medium components may act at a common point in the initiation of proliferation. We have evaluated the membrane transport of 86Rb, 3-O-methylglucose, AIB, and cycloleucine following stimulation of quiescent cells by Ca++ or serum. Serum stimulation results in large increases in the influx of all the substances tested. These increases are prevented if Ca++ is absent upon serum stimulation or they are rapidly diminished following Ca++ removal. In contrast, Ca++ stimulation of Ca++-deprived cells causes little or no enhancement of any of the transport systems, yet the cells progress to S phase in a manner similar to serum-stimulated cells. These results indicate that the Ca++ and serum G0 or G1 block are not equivalent and that the serum-induced change in transport of these components does not appear necessary for successful G1 phase progression. Furthermore, the data suggest that the sequence in which Ca++ or serum are presented to the cells alters the ability of Ca++ to modulate the transport systems. Quiescent cells which are exposed to Ca++ prior to serum possess a Ca++ modulation of several transport systems. Cells which are exposed to Ca++ subsequent to serum do not appear to possess this Ca++ regulation. 相似文献
58.
1. Protyrosinase from the egg of the grasshopper, Melanoplus differentialis, can be activated by excess sodium oleate or Aerosol. 2. The 3:4 quinone products of the reaction of activated protyrosinase with tyramine or tyrosine will oxidize ascorbic acid to dehydroascorbic acid. 3. The velocity of this latter oxidation of ascorbic acid increases with the amount of tyramine or tyrosine. 4. The oxidation of ascorbic acid by the tyramine-tyrosinase reaction delays the time of appearance of a red color associated with an indole quinone intermediary product in the formation of melanin. 5. Protyrosinase, in itself, and in the presence of tyrosinase substrates does not bring about the oxidation of ascorbic acid. 6. A naturally occurring substrate in a preparation of protyrosinase, sufficient to cause the oxidation of ascorbic acid, can be removed by dialysis against a 0.9 per cent sodium chloride solution. 7. Dialysis against such a solution does not change the properties of protyrosinase; the inactive enzyme must still be activated before it will catalyze the oxidation of tyramine or tyrosine. 8. When the natural substrate, tyrosine, or tyramine is absent, activation of protyrosinase does not result in the oxidation of ascorbic acid. 相似文献
59.
Antentor Hinton Jr. Prasanna Katti Margaret Mungai Duane D. Hall Olha Koval Jianqiang Shao Zer Vue Edgar Garza Lopez Rahmati Rostami Kit Neikirk Jessica Ponce Jennifer Streeter Brandon Schickling Serif Bacevac Chad Grueter Andrea Marshall Heather K. Beasley Young Do Koo Sue C. Bodine Nayeli G. Reyes Nava Anita M. Quintana Long-Sheng Song Isabella M. Grumbach Renata O. Pereira Brian Glancy E. Dale Abel 《Journal of cellular physiology》2024,239(4):e31281
60.
William J. Moore Jeffrey C. Kern Ramesh Bhat Peter V.N. Bodine Shoichi Fukyama Girija Krishnamurthy Ronald L. Magolda Keith Pitts Barb Stauffer Eugene J. Trybulski 《Bioorganic & medicinal chemistry》2010,18(1):190-201
Piperidinyl diphenylsulfonyl sulfonamides are a novel class of molecules that have inhibitory binding affinity for sFRP-1. As a secreted protein sFRP-1 inhibits the function of the secreted Wnt glycoprotein. Therefore, as inhibitors of sFRP-1 these small molecules facilitate the Wnt/β-catenin canonical signaling pathway. Details of the structure–activity relationships and biological activity of this structural class of compounds will be discussed. 相似文献