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41.
In vivo random mutagenesis of Bacillus subtilis by use of TnYLB-1, a mariner-based transposon 总被引:1,自引:0,他引:1
This report describes the construction and characterization of a mariner-based transposon system designed to be used in Bacillus subtilis, but potentially applicable to other gram-positive bacteria. Two pUC19-derived plasmids were created that contain the mariner-Himar1 transposase gene, modified for expression in B. subtilis, under the control of either sigmaA- or sigmaB-dependent promoters. Both plasmids also contain a transposable element (TnYLB-1) consisting of a Kan r cassette bracketed by the Himar1-recognized inverse terminal repeats, as well as the temperature-sensitive replicon and Erm r gene of pE194ts. TnYLB-1 transposes into the B. subtilis chromosome with high frequency (10(-2)) from either plasmid. Southern hybridization analyses of 15 transposants and sequence analyses of the insertion sites of 10 of these are consistent with random transposition, requiring only a "TA" dinucleotide as the essential target in the recipient DNA. Two hundred transposants screened for sporulation proficiency and auxotrophy yielded five Spo- clones, three with insertions in known sporulation genes (kinA, spoVT, and yqfD) and two in genes (ybaN and yubB) with unknown functions. Two auxotrophic mutants were identified among the 200 transposants, one with an insertion in lysA and another in a gene (yjzB) whose function is unknown. 相似文献
42.
Kashyap M Das D Preet R Mohapatra P Satapathy SR Siddharth S Kundu CN Guchhait SK 《Bioorganic & medicinal chemistry letters》2012,22(7):2474-2479
Scaffold hybridization of several natural and synthetic anticancer leads led to the consideration of indenoindolones as potential novel anticancer agents. A series of these compounds were prepared by a diversity-feasible synthetic method. They were found to possess anticancer activities with higher potency compared to etoposide and 5-fluorouracil in kidney cancer cells (HEK 293) and low toxicity to corresponding normal cells (Vero). They exerted apoptotic effect with blocking of cell cycle at G2/M phase. 相似文献
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Amrita Banerjee Arijit Jana Bikash R. Pati Keshab C. Mondal Pradeep K. Das Mohapatra 《The protein journal》2012,31(4):306-327
The tannase protein sequences of 149 bacteria and 36 fungi were retrieved from NCBI database. Among them only 77 bacterial
and 31 fungal tannase sequences were taken which have different amino acid compositions. These sequences were analysed for
different physical and chemical properties, superfamily search, multiple sequence alignment, phylogenetic tree construction
and motif finding to find out the functional motif and the evolutionary relationship among them. The superfamily search for
these tannase exposed the occurrence of proline iminopeptidase-like, biotin biosynthesis protein BioH, O-acetyltransferase,
carboxylesterase/thioesterase 1, carbon–carbon bond hydrolase, haloperoxidase, prolyl oligopeptidase, C-terminal domain and
mycobacterial antigens families and alpha/beta hydrolase superfamily. Some bacterial and fungal sequence showed similarity
with different families individually. The multiple sequence alignment of these tannase protein sequences showed conserved
regions at different stretches with maximum homology from amino acid residues 389–469 and 482–523 which could be used for
designing degenerate primers or probes specific for tannase producing bacterial and fungal species. Phylogenetic tree showed
two different clusters; one has only bacteria and another have both fungi and bacteria showing some relationship between these
different genera. Although in second cluster near about all fungal species were found together in a corner which indicates
the sequence level similarity among fungal genera. The distributions of fourteen motifs analysis revealed Motif 1 with a signature
amino acid sequence of 29 amino acids, i.e. GCSTGGREALKQAQRWPHDYDGIIANNPA, was uniformly observed in 83.3 % of studied tannase
sequences representing its participation with the structure and enzymatic function. 相似文献
46.
M. K. Pradhan L. Nayak P. N. Joshi P. K. Mohapatra L. Patro B. Biswal U. C. Biswal 《Photosynthetica》2008,46(3):370-377
Alterations in photosynthetic capacity of primary leaves of wheat seedlings in response to ultraviolet-B (UV-B; 280–320 nm;
60 μmol m−2 s−1) exposure alone and in combination with photosynthetically active radiation (PAR; 400–800 nm; 200 μmol m−2 s−1) during different phases of leaf growth and development were assessed. UV-B exposure resulted in a phase-dependent differential
loss in photosynthetic pigments, photochemical potential, photosystem 2 (PS2) quantum yield, and in vivo O2 evolution. UV-B exposure induced maximum damage to the photosynthetic apparatus during senescence phase of development. The
damages were partially alleviated when UV-B exposure was accompanied by PAR. UV-B induced an enhancement in accumulation of
flavonoids during all phases of development while it caused a decline in anthocyanin content during senescence. The differential
changes in these parameters demonstrated the adaptation ability of leaves to UV-B stress during all phases of development
and the ability was modified in UV-B+ PAR exposed samples. 相似文献
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This paper describes multiple shoot regeneration from leaf and nodal segments of a medicinally important herb Centella asiatica L. on Murashige and Skoog’s (MS) medium supplemented with a range of growth regulators. The highest number of multiple shoots
was observed on MS augmented with 3.0 mg dm−3 N6-benzylaminopurine (BAP) and 0.05 mg dm−3 α-naphthaleneacetic acid (NAA). Leaf explant showed maximum percentage of cultures regenerating shoots (81.6 %), with the
highest shoot number (8.3 shoots per explant) and the shoot length (2.1 cm) whereas, nodal explant showed less number of shoots
with callus formation at the base cut end. Successive shoot cultures were established by repeatedly sub-culturing the original
explant on a fresh medium. Rooting of in vitro raised shoots was best induced on half strength MS supplemented with 0.5 mg dm−3 indole-3-butyric acid (IBA) with highest percentage of shoot regenerating roots (76.8 %) with 3–4 roots per shoot. Plantlets
were acclimated in Vermi-compost and eventually established in soil. Contents of chlorophyll, total sugars, reducing sugars and proteins were estimated in
leaf tissue from both in vivo and in vitro raised plants. Chlorophyll content was higher in in vivo plants, whereas other three components were higher in in vitro plants. 相似文献
49.
Evaluation of maize microsatellite markers for genetic diversity analysis and fingerprinting in sugarcane. 总被引:4,自引:0,他引:4
The use of maize microsatellite markers as a potential cost-effective method for molecular analysis of sugarcane was evaluated. Of the 34 primer pairs obtained from maize genomic libraries, 14 showed repeatable amplifications in Saccharum species clones, commercial hybrids, and the related genera Erianthus, accounting for 41.17% cross transferability. Complex banding patterns were encountered in sugarcane with the number of amplified fragments ranging from 7 to 14 with an average of 10 per primer, indicating the high polyploidy and heterozygosity existing in sugarcane. Phenetic analysis of the SSR polymorphisms produced by nine primers could clearly differentiate the different species of Saccharum and Erianthus and revealed the relationships that existed between them. Genetic similarity co-efficient indicated low diversity existing among the S. officinarum clones (82%) and a relatively higher level of diversity in the S. spontaneum clones (69.7%). Higher level of divergence of Erianthus from Saccharum was also clearly estabilished. Five primers produced genus- and species-specific fragments for Erianthus, S. spontaneum, S. officinarum, and S. barberi. The polymorphic primers, when tested on a panel of 30 commercial sugarcane cultivars, revealed a broad range (32.4-83.3%) of pair-wise similarity values, indicating their ability to detect high levels of polymorphism. A combination of two primers could differentiate all the varieties, further emphasizing their potential in fingerprinting and varietal identification. 相似文献
50.