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The complete primary structure of the two hemoglobin components of the Great Indian Rhinoceros (Rhinoceros unicornis) is presented. The ratio for the two components B(alpha 2 beta I2): A(alpha 2 beta II2) is 6:4. Polypeptide subunits were separated by chromatography on CM-cellulose in a buffer containing 8M urea. The sequence was studied by degradation of the tryptic and hydrolytic cleavage products in a liquid phase sequencer. At position beta NA2 component B has Asp, whereas component A has Glu, an ATP-binding site in fish and reptilian hemoglobins. The other phosphate binding sites i.e. beta NA1 Val, beta EF6 Lys and beta H21 His are identical with 2,3-bisphosphoglycerate-(DPG)binding sites in mammalian hemoglobins, whereby rhinoceros hemoglobin resembles both ATP-sensitive poikilotherm hemoglobin and DPG-sensitive mammalian hemoglobin. The two components (beta I/beta II) additionally differ by exchange of Glu----Gly at position beta A3 and Gln----Lys at position beta GH3. The significance of these changes is discussed. Oxygenation properties of the two hemoglobins components and their dependence on ATP and DPG are given. The structure and function of Rhinoceros hemoglobin may give an insight into the evolution of the organic phosphate binding in vertebrate hemoglobins. 相似文献
4.
Summary Maize (Zea mays) leaf protoplasts were isolated from various leaves of two-week (4-leaf) seedlings and from sections of the third leaf blades. Microtubules (MTs) were visualized using immunofluorescence microscopy. Only freshly isolated protoplasts from the third and fourth leaf blades contained MTs, with protoplasts from the fourth leaf containing the most i.e. 13% of fourth-leaf protoplasts contained MTs. In general, protoplasts with fewer and smaller chloroplasts had more MTs. Initially 90–95% of protoplasts from basal portions of leaves had MTs but the percentage decreased slightly during culture particularly after 10 days. The antioxidant n-propyl gallate was beneficial in maintaining MT content. Few protoplasts from older sections intitially contained MTs but in all sections at least some protoplasts regained a significant MT content during culture (e.g., 10% of protoplast from the tip section possessed microtubules after 7 days of culture). Far fewer MTs were observed in individual leaf protoplasts than those isolated from suspension culture.Abbreviations BMS
Black Mexican Sweet
- MT
microtubule
- MtSB
microtubule stabilizing buffer
- PBS
phosphate buffered saline 相似文献
5.
DNA replication in maize leaf protoplasts 总被引:1,自引:0,他引:1
H. Wang Adrian J. Cutler M. Saleem Larry C. Fowke 《Plant Cell, Tissue and Organ Culture》1989,18(1):33-46
Maize leaf protoplasts were investigated for their metabolic competence and capacity to synthesize DNA. When protoplasts were incubated at elevated temperatures, they exhibited a heat shock response with specific proteins being preferentially synthesized. This indicated that the protoplasts were fully metabolically functional and capable of responding to environmental stimuli. Significant DNA synthesis was observed in these protoplasts after incorporation of 3H-thymidine into chromatin by trichloroacetic acid precipitation and by incorporation of 5-bromo-2-deoxyuridine (BrdU), an analog of thymidine, detected by immunofluorescence. The immunocytochemical method revealed that about 50% of nuclei in the maize leaf protoplasts were labelled after 3 days of culture and that most of these nuclei were labelled as intensely as normal mitotic cells. Aphidicolin, an inhibitor of DNA polymerase-, decreased the percentage of labelled nuclei, demonstrating that the labelling was substantially due to replicative DNA synthesis. However, chromosome condensation was not observed. It is proposed that these protoplasts are capable of DNA synthesis, but incapable of nuclear division. Effects of media additives on the number of nuclei entering S phase in these protoplasts were also assessed by the immunocytochemical method. Inclusion of 80mM Ca2+ in the enzyme solution increased protoplast yield and also appeared beneficial to DNA synthesis. The antioxidant, n-propyl gallate, which was used to stabilize the protoplasts, delayed the onset of DNA synthesis. Arginine and spermidine produced a slight increase in DNA synthesis.Abbreviations BrdU
5-bromo-2-deoxyuridine
- DMSO
dimethyl sulfoxide
- n-PG
n-propyl gallate
- PBS
phosphate-buffered saline
Dedicated to Dr. Friedrich Constabel on the occasion of his 60th birthday 相似文献
6.
Primary structure of -chain of pigeon is presented. It was determined by amino acid sequence analysis of intact -chain and its peptides obtained by the enzymatic and chemical cleavage. Comparison of amino acid sequence of the chain with other available data shows 14 Ile, 61 Lys, and 113 Ile as residues specific to pigeon. One important replacement at 11 contact is 55 MetSer. 相似文献
7.
The neem tree (Azadirachta indica) contains promising pest-control substances found effective against many economically important pests. These materials are easy to process by village-level industries and easy to use by limited-resource farmers, thereby offering potential for crop protection and off-farm income generation. The tree’s numerous pharmacological and other complementary uses make it doubly attractive for incorporation in large-scale rural development efforts. 相似文献
8.
The complete amino acid sequence of the major component of hemoglobin from amur-leopard (Panthera pardus orientalis) is presented. The major component accounts for more than 90% of the total hemoglobin. Separation of the globin subunits was achieved by ion-exchange chromatography on CM-cellulose in urea. The sequence was studied by automatic Edman degradation of tryptic and hydrolytic peptides. Alignment was carried out with human hemoglobin sequence. The NH2 terminus is blocked with Ac-serine. The data are compared with other mammalian hemoglobins and results are discussed with respect to sequence and physiology.85th communication on hemoglobin. 相似文献
9.
A highly specific proteolytic enzyme cleaving at the carboxyl group of valine has been isolated from Candida tropicalis. Its specificity has been determined by digesting beta-lactoglobulin and a number of synthetic peptides. The enzyme a glycoprotein, has a molecular mass of 40 +/- 7 kDa on the basis of sodium dodecyl sulphate polyacrylamide gel electrophoresis. Its optimum activity occurs at 37 degrees C at a pH between 8-9. It has been named "Valyl-proteinase" because of its selective cleavage. 相似文献
10.
Raman Kapur Mohammed Saleem Bryan L. Harvey Adrian J. Cutler 《In vitro cellular & developmental biology. Plant》1993,29(4):200-206
Summary Barley leaf blade protoplasts accumulate malonaldehyde, a product of lipid peroxidation, during culture. In addition, glutathione
levels fall after protoplast isolation and the proportion of glutathione in the oxidized state rises. These data indicate
oxidative stress after protoplast isolation and during culture. The cause of this phenomenon is revealed by data showing that
the activities of enzymes associated with antioxidative processes including glutathione reductase and ascorbate peroxidase
decrease after barley protoplast isolation. In contrast, protoplasts isolated from suspension cultured cells of bromegrass
and soybean exhibit little evidence for oxidative stress and increased activities of glutathione reductase and ascorbate peroxidase.
We suggest that an antioxidative response is associated with mitosis and colony formation from protoplasts, as exhibited by
bromegrass and soybean. Conversely, failure of an antioxidative response is associated with low viability and absence of mitosis,
as in barley. Increased viability of barley leaf protoplasts cultured on feeder layer cells is correlated with increased glutathione
content and higher glutathione reductase activity. 相似文献