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排序方式: 共有221条查询结果,搜索用时 15 毫秒
1.
Isolation and characterization of biologically active murine interleukin-1 alpha derived from expression of a synthetic gene in Escherichia coli 总被引:3,自引:0,他引:3
G O Daumy J M Merenda A S McColl G C Andrews A E Franke K F Geoghegan I G Otterness 《Biochimica et biophysica acta》1989,998(1):32-42
A murine interleukin-1 alpha (mIL-1 alpha) gene coding for amino acids 115 to 270 of the precursor protein (Lomedico, P.T., Gubler, U., Hellmann, C.P., Dukovich, M., Giri, J.G., Pan, Y.E., Collier, K., Semionow, R., Chua, A.O. and Mizel, S.B. (1984) Nature 312, 458-462) was chemically synthesized and expressed in Escherichia coli. mIL-1 alpha, in the form of insoluble inclusion bodies, accounted for approx. 30% of total cellular protein produced by the recombinant strain. A simple isolation protocol was developed in which inclusion body material was first solubilized in 3 M guanidine hydrochloride, and the mIL-1 alpha was then simultaneously purified and allowed to fold to its active conformation by dialysis against distilled water. This procedure yielded pure, biologically active mIL-1 alpha with 41% recovery of the mIL-1 alpha present in the guanidine hydrochloride extract. The purified preparation had the expected amino acid composition, a molar absorptivity of 28,200 M-1.cm-1 and a pI of 5.2. No methionyl-mIL-1 alpha was detected by N-terminal sequence analysis, and the endotoxin level was less than 10 pg per micrograms of mIL-1 alpha. The specific biological activity was 3.10(7) units/mg in a co-mitogenic thymocyte proliferation assay. In addition to full-length mIL-1 alpha, the preparation contained N-terminally truncated mIL-1 alpha species (mainly des-4 and des-6 amino acid forms). The truncated species were isolated and found to have the same biological activity as the complete polypeptide. Thus, the active fragment of mIL-1 alpha appears to consist of a proteinase-sensitive N-terminal region which is not essential for activity, and a proteinase-resistant core which harbors the essential determinants of its cytokine function. 相似文献
2.
The in-vitro efficacy of commercially available topical antimicrobial products against control strains and those from clinical material are compared with an agar diffusion model. The MICs of the constituent antimicrobial compounds have been determined for the same organisms. Plotting the inhibition zone diameters produced by the topical products against the log10 MICs of their constituent antimicrobial compound(s) gives overall product performance profiles for a range of organisms. These profiles confirm that the formulation of a topical product clearly modifies the response obtained with a specific antimicrobial compound. 相似文献
3.
4.
Penicillin G acylase from Proteus rettgeri is an 80,000- to 90,000-dalton enzyme composed of two nonidentical subunits. Both subunits were required for enzymatic activity. The 65,000-dalton beta subunit contained a phenylmethylsulfonyl fluoride-sensitive residue required for enzymatic activity, and the 24,500-dalton alpha subunit contained the domain that imparts specificity for the penicillin side chain. 相似文献
5.
Purification and characterization of 3-dehydroquinase from Escherichia coli. 总被引:1,自引:1,他引:0
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A procedure has been developed for the purification of 3-dehydroquinase from Escherichia coli. Homogeneous enzyme with specific activity 163 units/mg of protein was obtained in 19% overall yield. The subunit Mr estimated from polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate was 29,000. The native Mr, estimated by gel permeation chromatography on Sephacryl S-200 (superfine) and on TSK G3000SW, was in the range 52,000-58,000, indicating that the enzyme is dimeric. The catalytic properties of the enzyme have been determined and shown to be very similar to those of the biosynthetic 3-dehydroquinase component of the arom multifunctional enzyme of Neurospora crassa. 相似文献
6.
Nitella hookeri A. BR., a native charophyte of the Rotorua lakes in the North Island of New Zealand, was investigated in terms of its nutrient
requirements under laboratory culture in non-arenic conditions. Optimum growth in phosphate, nitrate, ammonia, iron and magnesium
occurred at 20 mg l−1 or lower. Higher concentrations of these ions were markedly inhibitory to growth when compared with the optimum. Calcium
and potassium gave growth at higher levels and over a wider range of concentrations. Sulphate and sulphur-amino acids do not
promote growth above the levels of the basal medium (1.2 mg l−1 sulphur as SO2−
4). Amino acids did not appear to provide a nitrogen source for the plant.
The optimum phosphate concentration was one thousand times higher than previously reported values. This may be due to the
presence of arsenic in the vegetative propagules used from Lake Rotoiti. Iron toxicity effects may have resulted from the
presence of EDTA.
The results are considered in relation to the known levels of nutrients present in lake waters. Only phosphorus appears directly
correlated with the natural distribution of the plant but it is suggested that anaerobic lake substrates may contribute iron
and ammonia to the plants and further explain their distribution. 相似文献
7.
Thylakoids and Photosystem II particles prepared from the cyanobacterium Synechococcus PCC 7942 washed with a HEPES/glycerol buffer exhibited low rates of light-induced oxygen evolution. Addition of either Ca2+ or Mg2+ to both thylakoids and Photosystem II particles increased oxygen evolution independently, maximal rates being obtained by addition of both ions. If either preparation was washed with NaCl, light induced O2 evolution was completely inhibited, but re-activated in the same manner by Ca2+ and Mg2+ but to a lower level. In the presence of Mg2+, the reactivation of O2 evolution by Ca2+ allowed sigmoid kinetics, implying co-operative binding. The results are interpreted as indicating that not only Ca2+, but also Mg2+, is essential for light-induced oxygen evolution in thylakoids and Photosystem II particles from Synechococcus PC 7942. The significance of the reactivation kinetics is discussed. Reactivation by Ca2+ was inhibited by antibodies to mammalian calmodulin, indicating that the binding site in Photosystem II may be analogous to that of this protein.Abbreviation HEPES
n-2-Hydroxyethylpiperazine--2-ethane sulphonic acid 相似文献
8.
mtDNA diversity in rhesus monkeys reveals overestimates of divergence time and paraphyly with neighboring species 总被引:4,自引:0,他引:4
Reconstructions of the human-African great ape phylogeny by using
mitochondrial DNA (mtDNA) have been subject to considerable debate. One
confounding factor may be the lack of data on intraspecific variation. To
test this hypothesis, we examined the effect of intraspecific mtDNA
diversity on the phylogenetic reconstruction of another Plio- Pleistocene
radiation of higher primates, the fascicularis group of macaque (Macaca)
monkey species. Fifteen endonucleases were used to identify 10 haplotypes
of 40-47 restriction sites in M. mulatta, which were compared with similar
data for the other members of this species group. Interpopulational,
intraspecific mtDNA diversity was large (0.5%- 4.5%), and estimates of
divergence time and branching order incorporating this variation were
substantially different from those based on single representatives of each
species. We conclude that intraspecific mtDNA diversity is substantial in
at least some primate species. Consequently, without prior information on
the extent of genetic diversity within a particular species, intraspecific
variation must be assessed and accounted for when reconstructing primate
phylogenies. Further, we question the reliability of hominoid mtDNA
phylogenies, based as they are on one or a few representatives of each
species, in an already depauperate superfamily of primates.
相似文献
9.
10.
Benzoate was established as the inducer of a unique 3-hydroxybenzoate 2-hydroxylase activity found in a Pseudomonas testosteroni mutant which is unable to grow on m-hydroxybenzoate as its sole source of carbon and energy. 相似文献