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21.
Type- and subcomplex-specific neutralizing antibodies against domain III of dengue virus type 2 envelope protein recognize adjacent epitopes 总被引:2,自引:1,他引:1
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Sukupolvi-Petty S Austin SK Purtha WE Oliphant T Nybakken GE Schlesinger JJ Roehrig JT Gromowski GD Barrett AD Fremont DH Diamond MS 《Journal of virology》2007,81(23):12816-12826
Neutralization of flaviviruses in vivo correlates with the development of an antibody response against the viral envelope (E) protein. Previous studies demonstrated that monoclonal antibodies (MAbs) against an epitope on the lateral ridge of domain III (DIII) of the West Nile virus (WNV) E protein strongly protect against infection in animals. Based on X-ray crystallography and sequence analysis, an analogous type-specific neutralizing epitope for individual serotypes of the related flavivirus dengue virus (DENV) was hypothesized. Using yeast surface display of DIII variants, we defined contact residues of a panel of type-specific, subcomplex-specific, and cross-reactive MAbs that recognize DIII of DENV type 2 (DENV-2) and have different neutralizing potentials. Type-specific MAbs with neutralizing activity against DENV-2 localized to a sequence-unique epitope on the lateral ridge of DIII, centered at the FG loop near residues E383 and P384, analogous in position to that observed with WNV-specific strongly neutralizing MAbs. Subcomplex-specific MAbs that bound some but not all DENV serotypes and neutralized DENV-2 infection recognized an adjacent epitope centered on the connecting A strand of DIII at residues K305, K307, and K310. In contrast, several MAbs that had poor neutralizing activity against DENV-2 and cross-reacted with all DENV serotypes and other flaviviruses recognized an epitope with residues in the AB loop of DIII, a conserved region that is predicted to have limited accessibility on the mature virion. Overall, our experiments define adjacent and structurally distinct epitopes on DIII of DENV-2 which elicit type-specific, subcomplex-specific, and cross-reactive antibodies with different neutralizing potentials. 相似文献
22.
Fossil preservation in the Burgess Shale 总被引:1,自引:0,他引:1
23.
Integrins are prominent receptors known from vertebrates and the higher
phyla of invertebrates. Until now, no evidence has been provided for the
existence of integrins in the lowest Metazoa, the sponges (Porifera). We
have isolated and characterized a cDNA clone encoding the alpha subunit of
integrin from the marine sponge Geodia cydonium (GCINTEG). The open reading
frame encodes a polypeptide of 1,086 residues (118 kDa). The intracellular
domain features the sequence Tyr- Phe-x-Gly-Phe-Phe-x-Arg, which is
different in one residue from the characteristic consensus pattern for
integrin alpha subunits. We conclude that sponges, the oldest multicellular
animal phylum, already utilize the structural elements which are required
for a tuned and controlled interaction among cells, and between cells and
the extracellular matrix.
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25.
Samples of grain from three spring barley cultivars of differing malting quality were collected at regular intervals during four weeks prior to harvest. The samples were dried, then assessed for relative grain hardness using the "Milling Energy" test. Ranking order of the cultivars for this character, which relates strongly to malting quality, was unaltered throughout. In a further experiment, it was demonstrated that selection for milling energy could be successfully practised on oven-dried grain collected six weeks after ear emergence. 相似文献
26.
TL Kieft EM Murphy DL Haldeman PS Amy BN Bjornstad EV McDonald DB Ringelberg DC White J Stair RP Griffiths TC Gsell WE Holben DR Boone 《Microbial ecology》1998,36(3):336-348
Abstract Two chronosequences of unsaturated, buried loess sediments, ranging in age from <10,000 years to >1 million years, were investigated to reconstruct patterns of microbial ecological succession that have occurred since sediment burial. The relative importance of microbial transport and survival to succession was inferred from sediment ages, porewater ages, patterns of abundance (measured by direct counts, counts of culturable cells, and total phospholipid fatty acids), activities (measured by radiotracer and enzyme assays), and community composition (measured by phospholipid fatty acid patterns and Biolog substrate usage). Core samples were collected at two sites 40 km apart in the Palouse region of eastern Washington State, near the towns of Washtucna and Winona. The Washtucna site was flooded multiple times during the Pleistocene by glacial outburst floods; the Winona site elevation is above flood stage. Sediments at the Washtucna site were collected from near surface to 14.9 m depth, where the sediment age was approximately 250 ka and the porewater age was 3700 years; sample intervals at the Winona site ranged from near surface to 38 m (sediment age: approximately 1 Ma; porewater age: 1200 years). Microbial abundance and activities declined with depth at both sites; however, even the deepest, oldest sediments showed evidence of viable microorganisms. Same-age sediments had equal quantities of microorganisms, but different community types. Differences in community makeup between the two sites can be attributed to differences in groundwater recharge and paleoflooding. Estimates of the microbial community age can be constrained by porewater and sediment ages. In the shallower sediments (<9 m at Washtucna, <12 m at Winona), the microbial communities are likely similar in age to the groundwater; thus, microbial succession has been influenced by recent transport of microorganisms from the surface. In the deeper sediments, the populations may be considerably older than the porewater ages, since microbial transport is severely restricted in unsaturated sediments. This is particularly true at the Winona site, which was never flooded. 相似文献
27.
Sulfation of fucoidin in focus embryos: III. Required for localization in the rhizoid wall
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Zygotes of the brown alga Fucus distichus L. Powell accumulate a sulfated polysaccharide (fucoidin) in the cell wall at the site of rhizoid formation. Previous work indicated that zygotes grown in seawater minus sulfate do not sulfate the preformed fucan (an unsulfated fucoidin) but form rhizoids. Under these conditions, we determined whether sulfation of the fucan is required for its localization in the rhizoid wall. This was accomplished by developing a specific stain for both the fucan and fucoidin. Using a precipitin assay, we demonstrated in vitro that the lectin ricin (RCA(I)) specifically complexes with both the sulfated and desulfated polysaccharide. No precipitate is observed when either is incubated in 0.1 M D-galactose or when RCA(I) is mixed with laminarin or alginic acid, the other major polysaccharides in Fucus. RCA(I) conjugated with fluorescein isothiocyanate (FITC) is also shown to bind specifically to fucoidin using a filter paper (DE81) assay. When added to zygotes, RCA(I)-FITC binds only to the site of fucoidin localization, i.e., the rhizoid cell wall. However, RCA(I)-FITC is not observed in the rhizoid wall of zygotes grown in the absence of sulfate. This observation is not due to inability of RCA(I)-FITC to bind to the fucan in vivo. Chemically desulfated cell walls that contained fucoidin in the rhizoid wall bind RCA(I)-FITC only in the rhizoid region. Also, the concentration of fucose-containing polymers and polysaccharides that form precipitates with RCA(I) is the same in embryos grown in the presence or absence of sulfate. If sulfate is added back to cultures of zygotes grown without sulfate, fucoidin is detected at the rhizoid tip by RCA(I)-FITC several hours later. These results support the conclusion that the enzymatic sulfation of the fucan is a modification of the polysaccharide required for its localization and/or assembly into a specific region of the cell wall. 相似文献
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29.
Lopez JV; Culver M; Stephens JC; Johnson WE; O'Brien SJ 《Molecular biology and evolution》1997,14(3):277-286
Differential rates of nucleotide substitution among different gene segments
and between distinct evolutionary lineages is well documented among
mitochondrial genes and is likely a consequence of locus-specific selective
constraints that delimit mutational divergence over evolutionary time. We
compared sequence variation of 18 homologous loci (15 coding genes and 3
parts of the control region) among 10 mammalian mitochondrial DNA genomes
which allowed us to describe different mitochondrial evolutionary patterns
and to produce an estimation of the relative order of gene divergence. The
relative rates of divergence of mitochondrial DNA genes in the family
Felidae were estimated by comparing their divergence from homologous
counterpart genes included in nuclear mitochondrial DNA (Numt, pronounced
"new might"), a genomic fossil that represents an ancient transfer of 7.9
kb of mitochondrial DNA to the nuclear genome of an ancestral species of
the domestic cat (Felis catus). Phylogenetic analyses of mitochondrial
(mtDNA) sequences with multiple outgroup species were conducted to date the
ancestral node common to the Numt and the cytoplasmic (Cymt) mtDNA genes
and to calibrate the rate of sequence divergence of mitochondrial genes
relative to nuclear homologous counterparts. By setting the fastest
substitution rate as strictly mutational, an empirical "selective
retardation index" is computed to quantify the sum of all constraints,
selective and otherwise, that limit sequence divergence of mitochondrial
gene sequences over time.
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30.
Loss of meiosis in Aspergillus 总被引:2,自引:0,他引:2
If strictly mitotic asexual fungi lack recombination, the conventional view
predicts that they are recent derivatives from older meiotic lineages. We
tested this by inferring phylogenetic relationships among closely related
meiotic and strictly mitotic taxa with Aspergillus conidial (mitotic)
states. Phylogenies were constructed by using DNA sequences from the
mitochondrial small ribosomal subunit, the nuclear ribosomal internal
transcribed spacers, and the nuclear 5.8S ribosomal gene. Over 920 bp of
sequence was analyzed for each taxon. Phylogenetic analysis of both the
mitochondrial and nuclear data sets showed at least four clades that
possess both meiotic and strictly mitotic taxa. These results support the
hypothesis that strictly mitotic lineages arise frequently from more
ancient meiotic lineages with Aspergillus conidial states. Many of the
strictly mitotic species examined retained characters that may be vestiges
of a meiotic state, including the production of sclerotia, sclerotium-like
structures, and hulle cells.
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