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1.
C M Mendel 《Biochimica et biophysica acta》1987,918(2):205-207
A number of studies conducted in the last decade showed that saturable ('specific') binding, by itself, does not necessarily imply biological significance. That is, biological ligands were shown to bind to inert materials as well as to biological receptors in a saturable manner. In these studies specific binding was operationally defined as binding that was displaceable by excess concentrations of unlabeled ligand. This method of measuring specific binding is now no longer considered optimal. To investigate whether optimal (computer-assisted) techniques of measuring specific binding--namely, nonlinear least-squares curve fitting of total binding data, with mathematical separation of the total binding into its various components--might ensure biological significance of measured specific binding, we studied the binding of high-density lipoproteins (HDL3) to tissue culture dishes as an example of binding without biological significance. This binding closely followed the paradigm of a ligand interacting with a class of homogeneous, saturable sites and with a class of relatively unsaturable sites, just as it would have if the HDL3 were interacting with an unpurified biological receptor. This finding indicates that computer-assisted analysis, while most accurately describing binding data, nevertheless does not ensure that measured specific binding has biological significance. Saturability is such a nonselective feature of equilibrium binding data that it should probably no longer be considered one of the criteria for deciding whether or not a defined binding site is a receptor. 相似文献
2.
A rapid method for cloning and sequencing variable-region genes of expressed immunoglobulins 总被引:9,自引:0,他引:9
A rapid procedure is described for cloning immunoglobulin V region genes from cells that express them. cDNA is synthesized from mRNA template using primers homologous to the immunoglobulin constant-region genes. Blunt-ended, double-stranded cDNA is obtained by sequential addition of enzymes to a single tube. The cDNA is inserted directly into the M13 vector, which is screened by plaque lifting for the presence of specific inserts. Screening probes can be generated from 32P-labeled single-stranded cDNAs generated from primers different from those used for cloning, or alternatively, from previously cloned V or C gene segments. The ease of cloning a cDNA V region is directly related to the abundance of Ig-specific mRNA within the cell of interest. This method minimizes the number of steps and the time needed to obtain accurate and complete sequences of any expressed Ig V region gene. 相似文献
3.
The activities of nitrite reductase (EC 1.7.7.1) are 60–70% of wild-type activity in pigment-deficient leaves of the chloroplast-ribosomedeficient mutants albostrians (Hordeum vulgare) and iojap (Zea mays). The activity and apoprotein of nitrate reductase (EC 1.6.6.1.) are lacking in the barley mutant. Only very low activities of nitrate reductase can be extracted from leaves of the maize mutant. The molybdenum cofactor of nitrate reductase and xanthine dehydrogenase (EC 1.2.3.2) is present in maize and barley mutant plants. However, it is not inducible by nitrate in pigment-deficient leaves of albostrians. From these results we conclude: (i) Nitrite reductase (a chloroplast enzyme) is synthesized in the cytoplasm and does not need the presence of nitrate reductase for the induction and maintenance if its activity. (ii) The loss or low activity of nitrate reductase is a consequence of the inability of the mutants to accumulate the apoprotein of this enzyme. (iii) The chloroplasts influence the accumulation (i.e. most probably the synthesis) of the nonchloroplast enzyme, nitrate reductase. The accumulation of nitrate reductase needs a chloroplast factor which is not provided by mutant plastids blocked at an early stage of their development.Abbreviations CRM
cross-reacting material
- Mo-co
molybdenum cofactor
- NiR
nitrite reductase
- NR
nitrate reductase 相似文献
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6.
The within tree distribution of some common natural enemies of bark beetles in pine plantations in Israel and some aspects
of their feeding habits were studied with special emphasis on the potential impact on the predatorAulonium ruficorne Olivier. A total of 12 predators and 2 parasite species were found associated withA. ruficorne in the natural enemy complex of bark beetles on pines. No secondary parasites were detected. The anthocoridScoloposcelis pulchella (Zetterstedt) and the dipteranMedetera striata Parent were observed feeding on immature stages ofA. ruficorne in the absence of scolytids. The associated Coleoptea:Nemosema elongatum F.,Rhizophagus bipustulatus L.,Corticeus spp.,Plastysoma spp. andPlegaderus discisus Erickson are thought to compete withA. ruficorne on larvae and pupae of bark beetles when the latter are in limited quantities (especially in the lower section of the tree).
The parasites, mainlyMetacolus unifasciatuss Forster andDendrosoter caenopachoides Ruschka are assumed to compete withA. ruficorne during the larval period in the smooth bark section of the stem. Competition might occur mainly during spring and fall. Deutonymphs
of the miteIpiduropoda sellnicki were detected on the abdomen ofA. ruficorne adults. Larvae of the predator were rarely infected in the field by the bacteriaSerratia sp. while laboratory cultures suffered high rate of mortality caused by this pathogen.
相似文献
7.
This paper describes a set of mathematical models to calculate runoff, soil erosion and chemical substance yields for drainage areas. The models represent different degrees of complexity and details of describing hydrological and hydrochemical processes. The availability of initial information and the requirements of the task determine the choice of a model for each particular case. These models have been written as PC/AT programs, to enable their use by a wide range of specialists. The models have been tested in different physical and geographical conditions, e.g. in some small watersheds on the Karelian Isthmus, west of Lake Ladoga, and in experimental catchment areas located in the Valdai Hills area in the southern part of Lake Ladoga drainage area. In further applications the drainage area models have been combined with models of the recipient water body, to obtain a single model of a watershed-lake system. The results of tests have proved that the models can be used for evaluation, simulation and forecasting of the processes of rainfall and snowmelt runoff, infiltration, evaporation, water erosion and pollutant wash-off from drainage areas. 相似文献
8.
Charles P. Moehs Paul V. Allen Mendel Friedman William R. Belknap 《Plant molecular biology》1996,32(3):447-452
We have isolated a cDNA encoding transaldolase, an enzyme of the pentose-phosphate pathway, from potato (Solanum tuberosum). The 1.5 kb cDNA encodes a protein of 438 amino acid residues with a molecular mass of 47.8 kDa. When the potato cDNA was expressed in Escherichia coli a 45 kDa protein with transaldolase activity was produced. The first 62 amino acids of the deduced amino acid sequence represent an apparent plastid transit sequence. While the potato transaldolase has considerable similarity to the enzyme from cyanobacteria and Mycobacterium leprae, similarity to the conserved transaldolase enzymes from humans, E. coli and Saccharomyces cerevisiae is more limited. Northern analysis indicated that the transaldolase mRNA accumulated in tubers in response to wounding. Probing the RNA from various potato tissues indicated that the transaldolase mRNA accumulation to higher levels in the stem of mature potato plants than in either leaves or tubers. These data are consistent with a role for this enzyme in lignin biosynthesis. 相似文献
9.
Cloning and expression of soluble epoxide hydrolase from potato 总被引:6,自引:1,他引:5
Andrew Stapleton Jeffrey K. Beetham Franck Pinot Joan E. Garbarino David R. Rockhold Mendel Friedman Bruce D. Hammock William R. Belknap 《The Plant journal : for cell and molecular biology》1994,6(2):251-258
Five cDNAs encoding a putative soluble epoxide hydrolase (sEH) from potato were isolated and characterized. The cDNAs contained open reading frames encoding 36 kDa polypeptides which were highly homologous to the carboxy terminal region of mammalian sEH. When one of the cDNAs was expressed in a baculovirus system a soluble 38 kDa protein with epoxide hydrolase activity was produced. The recombinant enzyme hydrolyzed a commonly used diagnostic substrate for the soluble form of mammalian EH. Inhibitor profiles of the recombinant potato and mammalian sEH were also similar. The expression of sEH in potato was found to be regulated by both developmental and environmental signals. Levels of mRNA for sEH were higher in meristematic tissue than in mature leaves. This mRNA was also observed to accumulate on wounding and application of exogenous methyl jasmonate. 相似文献
10.