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1.
2.
A R Thrower J Byrd E B Tarbet R K Mehra D H Hamer D R Winge 《The Journal of biological chemistry》1988,263(15):7037-7042
Metallothioneins have been isolated from Saccharomyces cerevisiae CUP1 mutants generated by Wright et al. (Wright, C. F., Hamer, D. H., and McKenney, K. (1986) Nucleic Acids Res. 14, 8489-8499). In the mutant metallothioneins, pairs of cysteinyl residues have been converted to seryl residues. The mutant proteins differ only in the positions of the double substitutions; each mutant molecule contains 10 cysteinyl residues. Each mutant protein lacks the first 8 residues at the amino terminus from the decoded gene sequence of the CUP1 locus. Mutant molecules consist of 53 residues analogous to the wild-type metallothionein and are designated 9/11, 24/26, 36/38, and 49/50 (in reference to the sequence positions of the Cys----Ser conversions). The properties of the mutant metallothioneins are vastly different, and host cells harboring the different plasmid-encoded mutant molecules show marked differences in sensitivity to CuSO4. Growth inhibition was observed at CuSO4 concentrations up to mM in cells containing the 9/11, 24/26, and 36/38 molecules, but not for cells containing protein 49/50. A CuSO4 concentration of 5 mM was required to inhibit the growth of yeast containing either 49/50 or the wild-type metallothionein. In the purified proteins the copper binding stoichiometry of each molecule, except protein 24/26, was nearly 8 mol eq. Protein 24/26 bound 5.5 copper ions/molecule. The Cu(I) chelator bathocuproine disulfonate reacted with over 50% of the copper ions in proteins 9/11, 24/26, and 36/38, but less than 10% of the copper ions in proteins 49/50 and wild-type metallothionein were reactive. The thiolates in 9/11, 24/26, and 36/38 were also more reactive in a disulfide exchange reaction with dithiodipyridine compared with the sulfhydryls in 49/50 and the wild-type molecules. The four mutant copper proteins are luminescent and exhibit a similar quantum yield. The cluster structures contributing to the particular electronic transitions are markedly more sensitive to oxygen in proteins 9/11, 24/26, and 36/38 compared with 49/50 and the wild-type molecules. The air-sensitive proteins exhibit a tertiary fold not recognized by polyclonal antibodies directed to a conformational epitope on yeast Cu-metallothionein. Protein 49/50 cross-reacts with the antibody in a concentration-dependent fashion similar to the wild-type protein. Mutation of 2 cysteinyl residues in the carboxyl portion of metallothionein does not significantly alter properties of the molecule, whereas mutation of several cysteines in the amino-terminal portion of the molecule yields a different conformation. 相似文献
3.
HLA and autoimmunity in North Indian type I (insulin-dependent) diabetic multiplex families 总被引:1,自引:0,他引:1
The HLA haplotype segregation and autoantibody spectrum in 7 type I (insulin-dependent) diabetic multiplex families of North Indian origin were determined. Of the total of 17 diabetic sibs, 7 shared both haplotypes and 3 shared one haplotype with the proband. No HLA-non-identical sibs were observed. This distribution of haplotypes was non-random (P approximately equal to 0.005). The mode of inheritance was compatible with an autosomal recessive model, while a dominant model was unlikely. Pancreatic islet-cell antibodies were found in 23.5% of affected sibs, but in no healthy family member. A high incidence of other autoantibodies (parietal-cell and thyroglobulin/thyroid microsomal antibodies) was detected in both the diabetic patients (26.3%), and in healthy first-degree relatives (22.2%). These findings emphasize the role of HLA-linked genes and autoimmunity in the pathogenesis of type I diabetes in North India. 相似文献
4.
Suppressor T lymphocytes from lepromatous leprosy skin lesions 总被引:13,自引:0,他引:13
R L Modlin V Mehra L Wong Y Fujimiya W C Chang D A Horwitz B R Bloom T H Rea P K Pattengale 《Journal of immunology (Baltimore, Md. : 1950)》1986,137(9):2831-2834
The immune response in leprosy forms a spectrum with lepromatous leprosy patients exhibiting specific unresponsiveness to antigens of Mycobacterium leprae. This unresponsiveness is thought to be related to the prevalence of T8-positive lymphocyte in these lepromatous lesions. To analyze the immunoregulatory function of these T8 cells, we developed simple procedures to extract lymphocytes from skin biopsy specimens of patients with leprosy. These lymphocytes were sorted for T8 and T4 positive cells, and cell lines were established by expansion with interleukin 2 (IL 2) and irradiated feeder cells. All T8 positive lines tested were positive for IL 2 receptors and HLA-DR determinants. These lines were additionally assayed for lepromin-induced suppression of the normal peripheral blood lymphocyte Con A proliferative response. Thirteen of 32 lines from six lepromatous patients showed significant suppressor activity, whereas nine lines from six tuberculoid patients and one line from normal peripheral blood failed to show suppression (p less than 0.001). Taken together, the finding of M. leprae-triggered suppressor cells within lepromatous skin lesions may in part explain the M. leprae unresponsiveness of lepromatous leprosy patients. 相似文献
5.
T. N. Khoshoo R. C. Mehra K. Bose 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1969,39(3):133-140
Summary
Ruellia tweediana and R. tuberosa are large flowered chasmogamous diploids (n=17) with normal meiosis and fertility. F
1 hybrids, successful in only one direction (R. tweediana x R. tuberosa), are vegetatively vigorous and possess 17 often heteromorphic bivalents with high degree of segregational irregularities. It is exclusively cleistogamous and completely pollen and seed sterile. Like F
1, the artificial amphidiploid (n=34) is also cleistogamous but shows preferential chromosome pairing with complete restoration of fertility. The parental chromosomes are sufficiently differentiated and cleistogamy is either genie or due to gene-cytoplasm interaction but sterility is entirely chromosomal. All floral parts excepting calyx are highly deformed. Such a deformity is associated with sterility in the F
1 but with fertility in the amphidiploid. This is perhaps the first case of origin by hybridization of a true breeding and fully fertile cleistogamous taxon from two chasmogamous species. It also shows the extent and nature of change in breeding system brought about by hybridization and/or polyploidy.The chromosome numbers in the six, out of 16, obligate cleistogamous taxa (Table 4) show that they are high polyploids. Perhaps their origin has been in the same manner as in the present case.
Zusammenfassung Ruellia tweediana und R. tuberosa sind großblütige, chasmogame Diploide (n=17) mit normaler Meiosis und Fertilität. Die F 1-Hybriden, die nur in einer Richtung gelingen (R. tweediana x R. tuberosa), sind vegetativ kräftig und besitzen häufig 17 heteromorphe Bivalente mit einem hohen Anteil an Spaltungsunregelmäßigkeiten. Die Hybride ist ausschließlich kleistogam und vollkommen pollen- und samensteril. Wie die F 1 ist auch die künstlich hergestellte Amphidiploide (n=34) kleistogam und zeigt eine präferentielle Chromosomenpaarung mit völliger Wiederherstellung der Fertilität. Die elterlichen Chromosomen sind genügend differenziert. Die Kleistogamie ist entweder genisch bedingt oder auf eine Gen-Cytoplasma-Interaktion zurückzuführen, die Sterilität ist ausschließlich durch die Chromosomen verursacht. Alle Teile der Blüte mit Ausnahme der Calyx sind stark deformiert. Bei der F 1 ist diese Deformation mit Sterilität verbunden, die amphidiploide Form ist jedoch fertil. Das ist vielleicht der erste Fall eines aus der Kreuzung zweier chasmogamer Spezies hervorgegangenen reinerbigen und voll fertilen kleistogamen Taxons. Es läßt sich auch der Umfang und die Art der durch Hybridisierung und durch Polyploidie verursachten Änderung des Zuchtsystems erkennen. Die Chromosomenzahl bei 6 von 16 obligaten kleistogamen Taxa (Tab. 4) zeigt, daß sie hochpolyploid sind. Vielleicht sind sie auf eine gleiche Weise wie im vorliegenden Falle entstanden.相似文献
6.
Purine hydroxylase II from Aspergillus nidulans has been purified to near homogeneity. The enzyme has a pI of 5.7, a molecular weight of 300,000, and two subunits with molecular weight of 153,000 each. The enzyme contains 2 FAD, 2 molybdenum atoms, and 4 (2 Fe-2S) iron-sulfur centers per molecule and exhibits broad specificity for reducing and oxidizing substrates. Among the more notable characteristics are the ability to oxidize hypoxanthine and nicotinic acid but not xanthine and virtually complete inactivity with oxygen. Moreover, while the enzyme is inactivated by borate and methanol, it is very resistant to cyanide and arsenite and it not inactivated by allopurinol. At infinite concentrations of reducing and oxidizing substrates, the Km for hypoxanthine was 119 microM, for nicotinic acid was 136 microM, and for NAD+ was 525 microM. 相似文献
7.
Species differences in the occurrence of copper-metallothionein in the particulate fractions of the liver of copper-loaded animals. 总被引:1,自引:1,他引:0 下载免费PDF全文
Large amounts of Cu-metallothionein were obtained by 2-mercaptoethanol and sodium dodecyl sulphate extractions of the particulate fractions of the liver of pigs given high-Cu2+ diets or rats injected with Cu2+. Three isoproteins were purified from pig liver and characterized on the basis of their physicochemical properties, metal content and amino acid composition. No such pool of Cu-metallothionein was present in the liver of Cu2+-loaded sheep or of rats given Cu2+-supplemented diets. 相似文献
8.
Biochemical and genetic characterization of three hamster cell mutants resistant to diphtheria toxin 总被引:2,自引:0,他引:2 下载免费PDF全文
RK Draper D Chin D Eurey-Owens IE Scheffler MI Simon 《The Journal of cell biology》1979,83(1):116-125
We describe here three different hamster cell mutants which are resistant to diphtheria toxin and which provide models for investigating some of the functions required by the toxin inactivates elongation factor 2 (EF-2). Cell-free extracts from mutants Dtx(r)-3 was codominant. The evidence suggests that the codominant phenotype is the result of a mutation in a gene coding for EF-2. The recessive phenotype might arise by alteration of an enzyme which modifies the structure of EF-2 so that it becomes a substrate for reaction with the toxin. Another mutant, Dtx(r)-2, contained EF-2 that was sensitive to the toxin and this phenotype was recessive. Pseudomonas aeruginosa exotoxin is known to inactivate EF-2 as does diphtheria toxin and we tested the mutants for cross-resistance to pseudomonas exotoxin. Dtx(r)-1 and Dtx(r)-3 were cross-resistant while Dtx(r)-2 was not. It is known that diphtheria toxin does not penetrate to the cytoplasm of mouse cells and that these cell have a naturally occurring phenotype of diphtheria toxin resistance. We fused each of the mutants with mouse 3T3 cells and measured the resistance. We fused each of the mutants with mouse 3T3 cells and measured the resistance of the hybrid cells to diphtheria toxin. Intraspecies hybrids containing the genome of mutants Dtx(r)-1 and Dtx(r)-3 had some resistance while those formed with Dtx(r)-2 were as sensitive as hybrids derived from fusions between wild-type hamster cells and mouse 3T3 cells. 相似文献
9.
Approximately 43–60% of the total genome in bovine, goat and sheep consisted of interspersed repeated and single copy DNA
sequences. Most of the interspersed repeated DNA sequences were 1500–2400 nucleotide pair long while a minor portion was more
than 4000 nucleotide pair long in goat and sheep and 3200 nucleotide pair long in bovine. About 1/3rd of single copy sequence
were interspersed and their length was in the range of 1000–1500 nucleotide pairs. 相似文献
10.