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111.
Cloning, expression, purification, and characterization of the acid alpha-mannosidase from Trypanosoma cruzi 总被引:1,自引:0,他引:1
Vandersall-Nairn AS; Merkle RK; O'Brien K; Oeltmann TN; Moremen KW 《Glycobiology》1998,8(12):1183-1194
The acid alpha-mannosidase of Trypanosoma cruzi is a broad-specificity
hydrolase involved in the catabolism of glycoconjugates, presumably in the
digestive vacuole. We have cloned the alpha-mannosidase gene from a T.cruzi
epimastigote genomic library. The alpha-mannosidase gene was determined to
be single copy by Southern analysis, and similar sequences were not
detected in genomic digests of either Trypanosoma brucei or Leishmania
donovani. The coding region was subcloned into the Pichia pastoris
expression vector pPICZ, and alpha-mannosidase activity was detected in the
medium of induced cultures. The recombinant alpha- mannosidase demonstrated
a pH optimum, inhibition by swainsonine, Km, and substrate specificity
consistent with the characteristics of the alpha-mannosidase previously
purified from T.cruzi epimastigotes. The recombinant enzyme was purified
103-fold from the culture medium of Pichia pastoris and had a native
molecular mass of 359 kDa by gel filtration. A combination of SDS-PAGE,
deglycosylation with endo H, and NH2-terminal sequencing indicates that the
enzyme is originally synthesized as a homodimeric polypeptide that is
subsequently cleaved to form a heterotetramer composed of 57 and 46 kDa
subunits. A polyclonal antibody raised to the recombinant enzyme was shown
to immunoprecipitate the alpha-mannosidase from T.cruzi cell extracts and
will be used in future immunolocalization studies.
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112.
The development of antimicrobial drug resistance has encouraged scientists to develop alternate methods to combat infectious pathogens associated with dental diseases. Therefore, it is of interest to predict interactions for catechin (a plant derived compound) with protein targets in the red complex pathogens using computer aided network tools. However, in vitro and in vivo studies are warranted to confirm the antimicrobial effect of catechin (gallocatechin, epicatechin, epigallactocatechin (EGC) and gallolyl catechins) on the dental pathogens. 相似文献
113.
Background
Colorants derived from natural sources look set to overtake synthetic colorants in market value as manufacturers continue to meet the rising demand for clean label ingredients – particularly in food applications. Many ascomycetous fungi naturally synthesize and secrete pigments and thus provide readily available additional and/or alternative sources of natural colorants that are independent of agro-climatic conditions. With an appropriately selected fungus; using in particular chemotaxonomy as a guide, the fungal natural colorants could be produced in high yields by using the optimized cultivation technology. This approach could secure efficient production of pigments avoiding use of genetic manipulation. 相似文献114.
Andy?Pang Andrew?D?Smith Paulo?AS?Nuin Elisabeth?RM?TillierEmail author 《BMC bioinformatics》2005,6(1):236
Background
General protein evolution models help determine the baseline expectations for the evolution of sequences, and they have been extensively useful in sequence analysis and for the computer simulation of artificial sequence data sets. 相似文献115.
116.
Destruction of lysine by nonreducing sugars 总被引:1,自引:0,他引:1
117.
A new method for the enrichment of cultures of Serratia marcescens for auxotrophic mutants has been developed. The method is based on the formation of filaments by growing cells in minimal medium M70 containing azthreonam. Auxotrophic mutants unable to grow in M70 do not form filaments. Mutants are collected from the culture by filtration. 相似文献
118.
119.
120.
Evolutionary history of the COII/tRNALys intergenic 9 base pair deletion in human mitochondrial DNAs from the Pacific 总被引:14,自引:2,他引:12
Redd AJ; Takezaki N; Sherry ST; McGarvey ST; Sofro AS; Stoneking M 《Molecular biology and evolution》1995,12(4):604-615
Length changes in human mitochondrial DNA (mtDNA) are potentially useful
markers for inferring the evolutionary history of populations. One such
length change is a nine base pair (9-bp) deletion that is located in the
intergenic region between the COII gene and the Lysine tRNA gene
(COII/tRNALys intergenic region). This deletion has been used as a genetic
marker to trace descent from peoples of East Asian origin. A geographic
cline of the deletion frequency across modern Pacific Islander populations
suggests that the deletion may be useful for tracing prehistoric Polynesian
origins and affinities. Mitochondrial DNA sequence variation within two
variable segments of the control region (CR) permits a number of inferences
regarding the evolutionary history of the 9-bp deletion that cannot be
determined from frequency data alone. We obtained CR sequences from 74
mtDNAs with the 9-bp deletion from Indonesia, coastal Papua New Guinea
(PNG), and American Samoa. Phylogenetic and pairwise distribution analysis
of these CR sequences pooled with previously published CR sequences reveals
that the deletion arose independently in Africa and Asia and suggests
possible multiple origins of the deletion in Asia. A clinal increase of the
frequency of the 9-bp deletion across the three Pacific populations is
associated with a decrease in CR sequence diversity, consistent with
founder events. Furthermore, analysis of pairwise difference distributions
indicates an expansion time of proto-Polynesians that began 5,500 yr ago
from Southeast Asia. These results are consistent with the express train
model of Polynesian origins.
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