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1.
DNA fragments containing theKlebsiella oxytoca genes encoding -glucosidase and amylase were cloned into the kanamycin resistance transposon Tn5. Another DNA fragment containing two genes for polygalacturonatetrans-eliminase was cloned into Tn1721. These newly constructed transposons were then each transposed in vivo onto the broad-host-range plasmid pR751 and conjugally transferred to a variety of Gram-negative bacteria. These were then screened for the newly acquired phenotypes. 相似文献
2.
Enhanced recognition of a modified peptide antigen by cytotoxic T cells specific for influenza nucleoprotein 总被引:10,自引:0,他引:10
A previously identified Kd restricted epitope of influenza A virus nucleoprotein (147-161) was modified, resulting in recognition by Kd restricted cytotoxic T cells at significantly lower concentrations than the natural peptide sequence. This was achieved by first refining the epitope to the minimum determinant 147-158. Deletion of arginine 156 resulted in a peptide that was shown to be greatly superior in both dose response titrations and in its rate of association with cells to form targets. Analog peptides were tested to determine the important amino acid changes. These data suggest that T cell epitopes can be modified to result in improved immunological recognition. 相似文献
3.
4.
Genetic and physical map of the 2,4-dichlorophenoxyacetic acid-degradative plasmid pJP4. 总被引:29,自引:12,他引:17
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Plasmid pJP4 is an 80-kilobase, IncP1, broad-host-range conjugative plasmid of Alcaligenes eutrophus encoding resistance to mercuric chloride and phenyl mercury acetate and degradation of 2,4-dichlorophenoxyacetic acid, 2-methyl-4-chlorophenoxyacetic acid, and 3-chlorobenzoate. By the use of cloning, transposon mutagenesis, and restriction endonuclease analysis, a biophysical and genetic map of pJP4 was generated. 相似文献
5.
Plasmid pULB113 (RP4::Mini-Mu) promoted homologous gene transfer inAeromonas hydrophila; transfer of chromosomal markers occurred at frequencies of between 10–3 and 10–4 per donor cell regardless of the marker selected; this indicated chromosome transfer from multiple origins. With a variety of amino acid biosynthetic markers, a single circular map of this bacterium was constructed. 相似文献
6.
Maintenance of the cellobiose utilization genes of Escherichia coli in a cryptic state 总被引:6,自引:1,他引:5
The genes for cellobiose utilization are normally cryptic in Escherichia
coli. The cellobiose system was used as a model to understand the process
by which silent genes are maintained in microbial populations. Previously
reported was (1) the isolation of a mutant strain that expresses the
cellobiose-utilization (Cel) genes and (2) that expression of those genes
allows utilization of three beta- glucoside sugars: cellobiose, arbutin,
and salicin. The Cel gene cluster has now been cloned from that mutant
strain. In the course of locating the Cel genes within the cloned DNA
segment, it was discovered that inactivation of the Cel-encoded hydrolase
rendered the host strain sensitive to all three beta-glucosides as potent
inhibitors. This sensitivity arises from the accumulation of the
phosphorylated beta- glucosides. Because even the fully active genes
conferred some degree of beta-glucoside sensitivity, the effects of
cellobiose on a series of five Cel+ mutants of independent origin were
investigated. Although each of those strains utilizes cellobiose as a sole
carbon and energy source, cellobiose also acts as a potent inhibitor that
reduces the growth rate on glycerol 2.5-16.5-fold. On the other hand,
wild-type strains that cannot utilize cellobiose are not inhibited. The
observation that the same compound can serve either as a nutrient or as an
inhibitor suggests that, under most conditions in which cellobiose will be
present together with other resources, there is a strong selective
advantage to having the cryptic (Cel0) allele. In those environments in
which cellobiose is the sole, or the best, resource, mutants that express
the genes (Cel+) will have a strong selective advantage. It is suggested
that temporal alternation between these two conditions is a major factor in
the maintenance of these genes in E. coli populations. This alternation of
environments and fitnesses was predicted by the model for cryptic-gene
maintenance that was previously published.
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7.
Stable albicidin resistance in Escherichia coli involves an altered outer-membrane nucleoside uptake system 总被引:2,自引:0,他引:2
Albicidin blocked DNA synthesis in intact cells of a PolA- EndA- Escherichia coli strain, and in permeabilized cells supplied with all necessary precursor nucleotides, indicating a direct effect on prokaryote DNA replication. Replication of phages T4 and T7 was also blocked by albicidin in albicidin-sensitive (Albs) but not in albicidin-resistant (Albr) E. coli host-cells. All stable spontaneous Albr mutants of E. coli simultaneously became resistant to phage T6. The locus determining albicidin sensitivity mapped at tsx, the structural gene for an outer-membrane protein used as a receptor by phage T6 and involved in transport through the outer membrane of nucleosides present at submicromolar extracellular concentrations. Albicidin does not closely resemble a nucleoside in structure. However, Albs E. coli strains rapidly accumulated both nucleosides and albicidin from the surrounding medium whereas the Albr mutants were defective in uptake of nucleosides and albicidin at low extracellular concentrations. An insertion mutation blocking Tsx protein production also blocked albicidin uptake and conveyed albicidin resistance. Albicidin supplied at approximately 0.1 microM blocked DNA replication within seconds in intact Albs E. coli cells, but a 100-fold higher albicidin concentration was necessary for a rapid inhibition of DNA replication in permeabilized cells. We conclude that albicidin is effective at very low concentrations against E. coli because it is rapidly concentrated within cells by illicit transport through the tsx-encoded outer-membrane channel normally involved in nucleoside uptake. Albicidin resistance results from loss of the mechanism of albicidin transport through the outer membrane. 相似文献
8.
J. M. Pemberton S. D. Albon F. E. Guinness T. H. Clutton-Brock R. J. Berry 《Evolution; international journal of organic evolution》1988,42(5):921-934
The survival of red deer (Cervus elaphus L.) calves to two years of age was examined in relation to electrophoretic variation in a population on the Scottish island of Rhum. Survival was analyzed using logistic analysis in which the “phenotypic” factors birth weight, birth date, subdivision of the study area, cohort, and sex, which affect the probability of a calf's survival, were taken into account. All three polymorphic loci examined, Mpi, Idh-2, and Trf (each with two detected alleles) are significantly associated with juvenile survival. At Mpi, there is selection against one allele, f (or an allele at a linked locus), and there are indications that this effect is stronger in females than males. For Idh-2, overall, the heterozygote class survives better than the two homozygotes, which survive equally well. However, again there is a difference between the sexes; female heterozygotes survive much better than homozygotes, whereas male homozygotes survive better than heterozygotes, and the difference in survival is smaller. Furthermore, there is an interaction involving Mpi, Idh-2, and survival in which Mpif carriers that are also Idh-2 homozygotes survive very badly compared with other Mpi-Idh-2 combinations, which all survive equally well. For Trf, the heterozygote class survives best, and there is also a difference in survival between the two homozygote classes. Genotype frequencies in the adult population are consistent with the results for calf survival, in that the Mpif frequency is lower in succeeding cohorts of surviving adults, whereas no significant gene frequency change is apparent for Idh-2 or Trf. 相似文献
9.
T. Komiyama H. Grn P. A. Pemberton G. S. Salvesen 《Protein science : a publication of the Protein Society》1996,5(5):874-882
Serpins are well-characterized inhibitors of the chymotrypsin family serine proteinases. We have investigated the interaction of two serpins with members of the subtilisin family, proteinases that possess a similar catalytic mechanism to the chymotrypsins, but a totally different scaffold. We demonstrate that alpha 1 proteinase inhibitor inhibits subtilisin Carlsberg and proteinase K, and alpha 1 antichymotrypsin inhibits proteinase K, but not subtilisin Carlsberg. When inhibition occurs, the rate of formation and stability of the complexes are similar to those formed between serpins and chymotrypsin family members. However, inhibition of subtilisins is characterized by large partition ratios where more than four molecules of each serpin are required to inhibit one subtilisin molecule. The partition ratio is caused by the serpins acting as substrates or inhibitors. The ratio decreases as temperature is elevated in the range 0-45 degrees C, indicating that the serpins are more efficient inhibitors at high temperature. These aspects of the subtilisin interaction are all observed during inhibition of chymotrypsin family members by serpins, indicating that serpins accomplish inhibition of these two distinct proteinase families by the same mechanism. 相似文献
10.
Viral R plasmid Rphi6P: properties of the penicillinase plasmid prophage and the supercoiled, circular encapsidated genome.
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Properties of the viral R plasmid Rphi6P are described. As a temperate bacteriophage, it plaques on the facultative phototroph Rhodopseudomonas sphaeroides. Under aerobic conditions the phage had a latent period of 180 min, a burst time of 200 min, and a burst size of 15 to 20 particles per infective center. The encapsidated viral genome occurred as a supercoiled, circular DNA duplex with a mean contour length of 16.5 +/- 10 micron. Percent guanine plus cytosine, as calculated from thermal denaturation profiles, was 63.5. Mitomycin C-induced loss of the prophage suggested an extrachromosomal location in the host cell. Use of this curing agent enabled the isolation of a plasmid-free strain of R. sphaeroides. Biophysical analysis of the plasmid-free strain lysogenized with Rphi6P confirmed that the prophage occurred as a plasmid in the host cell. 相似文献