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61.
2-Chloroacetaldehyde and 2-chloroacetal are potent inhibitors of DNA synthesis in animal cells 总被引:2,自引:0,他引:2
J C Kandala J E Mrema A DeAngelo F B Daniel R V Guntaka 《Biochemical and biophysical research communications》1990,167(2):457-463
The effect of 2-chloroacetaldehyde, CAA, a metabolite of vinyl chloride and 2-chloroacetal, CAC, an ethyl diester of chloroacetaldehyde, on DNA synthesis in animal cells has been investigated. Both compounds drastically inhibited DNA synthesis at 10 to 20 microM. The inhibitory effect of the chemicals appears to be directly on DNA synthesis rather than on the uptake of thymidine or the formation of nucleotides. Residual DNA made in the presence of CAA had an average chain length of 300 nucleotides compared to a length of several thousand nucleotides in the absence of CAA. Synchronization experiments revealed that the inhibitory effect is reversible if 2-chloroacetaldehyde is removed within two hours but not after longer exposures. 相似文献
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Despite major advances in characterizing purine(R)-purine(R), purine(R)-pyrimidine(Y) and pyrimidine(Y)-pyrimidine(Y) mismatches in DNA, there have not been any structural studies on a synthetic DNA duplex containing several different mispairs. Here, using NMR restrained molecular mechanics and dynamics simulations we have structurally characterized a 12 nucleotide long antiparallel DNA duplex with three different mispairs, namely A+-C, G-T and T-C. Our results show that the overall conformation of the antiparallel DNA duplex is B-DNA-like with slight structural distortions at or near the mispairs' sites. All these mispairs are properly stacked with their flanking base pairs. Each mispair is stabilized by two hydrogen bonds and the decreasing order of the hydrogen-bonding interactions is G-T>T-C>A+-C. G-T mispair has smaller configurational space while the structure is slightly bent at A+-C mispair's site. Overall, this study is the first ever structural characterization of a DNA duplex with three different mismatched base pairs and throws light upon the local conformations of the three mispairs present in the DNA duplex. 相似文献
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Ramanathan S Chary KV Rao BJ 《Biochemical and biophysical research communications》2002,295(3):730-736
G-G base-paired hairpin DNA structures on template strands offer potential "road-blocks" to a traversing polymerase. Klenow polymerase (exo+) pauses while replicating through G-G base-paired hairpin DNA due to the generation of G-G:C triplex. However, exonuclease-deficient Klenow traverses through de novo generated G-G:C triplexes leading to full-length C:G duplexes. Alleviation of such road-blocks by exo- Klenow ensues faster at lower Mg2+, a kinetic effect consistent with the role of Mg2+ in stabilizing G-G:C triplex fold. The ability of exonuclease-deficient polymerase to go past the de novo generated G-G:C triplexes suggests that the "idling" of exo+ polymerase at G-G road-block is due to the reiterative polymerase/exonuclease action. The full-length replication product carrying a C(n)-G(n) duplex at one end is further "expanded" by exo- Klenow through C-strand "slippage" leading to the generation of C+-G:C triplex, which is exemplified by the premature arrest of the same at low pH that further stabilizes the C+-G:C triplex. 相似文献
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Venkatraman Ramanujam Sunita Patel Atul K. Srivastava Yogendra Sharma Kandala V. R. Chary 《Biomolecular NMR assignments》2013,7(2):221-224
The sequence specific backbone 1H, 13C and 15N resonance assignments of an intrinsically unstructured βγ-crystallin from Hahella chejuensis are reported. The secondary structure chracterization of the unstructured protein reveals that large fraction of residues exhibits β-strand propensity, as in the case of the Ca2+-bound structured protein. 相似文献
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The 6-kDa early secretory antigenic target (ESAT-6) and culture filtrate protein-10 (CFP-10), expressed from the region of deletion-1 (RD1) of Mycobacterium tuberculosis H37Rv, are known to play a key role in virulence. In this study, we explored the thermodynamic and biochemical changes associated with the formation of the 1 : 1 heterodimeric complex between ESAT-6 and CFP-10. Using isothermal titration calorimetry (ITC), we precisely determined the association constant and free energy change for formation of the complex to be 2 x 10(7) M(-1) and -9.95 kcal.mol(-1), respectively. Strikingly, the thermal unfolding of the ESAT-6-CFP-10 heterodimeric complex was completely reversible, with a T(m) of 53.4 degrees C and DeltaH of 69 kcal.mol(-1). Mixing of ESAT-6 and CFP-10 at any temperature below the T(m) of the complex led to induction of helical conformation, suggesting molecular recognition between specific segments of unfolded ESAT-6 and CFP-10. Enhanced biochemical stability of the complex was indicated by protection of ESAT-6 and an N-terminal fragment of CFP-10 from proteolysis with trypsin. However, the flexible C-terminal of CFP-10 in the complex, which has been shown to be responsible for binding to macrophages and monocytes, was cleaved by trypsin. In the presence of phospholipid membranes, ESAT-6, but not CFP-10 and the complex, showed an increase in alpha-helical content and enhanced thermal stability. Overall, complex formation resulted in structural changes, enhanced thermodynamic and biochemical stability, and loss of binding to phospholipid membranes. These features of complex formation probably determine the physiological role of ESAT-6, CFP-10 and/or the complex in vivo. The ITC and thermal unfolding approach described in this study can readily be applied to characterization of the 11 other pairs of ESAT-6 family proteins and for screening ESAT-6 and CFP-10 mutants. 相似文献
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Ruth Pulikottil-Jacob Martin Connock Ngianga-Bakwin Kandala Hema Mistry Amy Grove Karoline Freeman Matthew Costa Paul Sutcliffe Aileen Clarke 《PloS one》2016,11(11)
BackgroundTotal hip replacement for end stage arthritis of the hip is currently the most common elective surgical procedure. In 2007 about 7.5% of UK implants were metal-on-metal joint resurfacing (MoM RS) procedures. Due to poor revision performance and concerns about metal debris, the use of RS had declined by 2012 to about a 1% share of UK hip procedures. This study estimated the lifetime cost-effectiveness of metal-on-metal resurfacing (RS) procedures versus commonly employed total hip replacement (THR) methods.Conclusion/SignificanceOur results imply that in most cases RS has not been a cost-effective resource and should probably not be adopted by decision makers concerned with the cost effectiveness of hip replacement, or by patients concerned about the likelihood of revision, regardless of patient age or gender. 相似文献
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