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1.
J Kwang S Bolin E T Littledike E J Dubovi R O Donis 《Biochemical and biophysical research communications》1991,178(3):1326-1334
A 917-base pair segment of the bovine viral diarrhea virus (BVDV) genome encoding part of the p80 region was cloned into plasmid Gex-2T expression vector for expression as a fusion protein with glutathione-S-transferase (GST). When the p80 and GST sequences were in the same reading frames, the resulting GST-p80 fusion protein had a molecular mass of 58 kilodaltons (kDa) in SDS-PAGE. Extracts of control E. coli carrying only the vector plasmid (Gex-2T) did not contain this new 58-kDa protein band. Mouse monoclonal antibody specific to BVDV-p80 recognized this recombinant protein. Seventy cattle sera that had an SN titer (to TGAC isolate of cytopathic BVDV) greater than 1:8 reacted with this recombinant protein in Western blots. Of 28 cattle sera that had SN titers less than or equal to 1:8, only one serum tested positive on Western blots. 相似文献
2.
S N Walters T A Reinhardt M A Dominick R L Horst E T Littledike 《Archives of biochemistry and biophysics》1986,246(1):366-373
In order to investigate the subcellular distribution of unoccupied 1,25-dihydroxyvitamin D3 receptors, highly purified cytoplasts and nucleoplasts were prepared from two kidney cell lines (PK1 and MDBK). This was accomplished utilizing the technique of enucleation by cytochalasin B and density gradient centrifugation. Unoccupied 1,25-dihydroxyvitamin D3 receptors were found in both the nuclear and cytosolic compartments, with approximately 70% of the receptors localized in the cytoplasm. When cells were pretreated with 1,25-[3H]dihydroxyvitamin D, prior to enucleation, it was found that 90% of the receptor-hormone complex was associated with nucleoplasts, thus demonstrating that cytochalasin B treatment does not alter the high-affinity association of the receptor-hormone complex with the nucleus. The ratio of unoccupied receptor/protein was found to be the same in whole cells, cytoplasts, and nucleoplasts for both cell types. The ratio of unoccupied receptor/DNA was highest in cytoplasts and lowest in nucleoplasts. Taken together, these data indicate that the unoccupied 1,25-dihydroxyvitamin D receptor is generally associated with cell proteins and not specifically associated with cell DNA. We therefore propose, at least for these cells, that the unoccupied 1,25-dihydroxyvitamin D receptor exists in equilibrium between the nuclear and cytosolic compartments of the whole cell, and receptor-hormone binding shifts this equilibrium to favor nuclear localization. 相似文献
3.
Timothy A. Reinhardt Ronald L. Horst E.Travis Littledike Donald C. Beitz 《Biochemical and biophysical research communications》1982,106(3):1012-1018
1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] receptor was characterized after partial purification of thymus cytosol by ammonium sulfate fractionation. The 1,25-(OH)2D3 receptor sediments at 3.7S in 5–20% sucrose gradients. The binding of 1,25-(OH)2D3 in thymic cytosol was a saturable process with high affinity (Kd = 0.12?0.48 nM) at 4°C. Competition for 1,25-(OH)2[3H]D3 receptor by nonradioactive analogs demonstrated the affinities of these analogs to be in order; 1,25-(OH)2D3 = 1,24R,25-(OH)3D3 = 1,25S,26-(OH)3D3 = 1,25R,26-(OH)3D3 > 1,25-(OH)2D3-26,23 lactone > 25-OHD3 > 23R,25-(OH)2D3 > 24R,25-(OH)2D3 > 23S,25-(OH)2D3 ? 25-OHD3-26,23 lactone. The receptor bound to DNA cellulose columns in low salt buffer and eluted as a single peak at 0.21 M KCl. These findings provide evidence that the thymus possesses a 1,25-(OH)2D3 receptor with properties indistinguishable from 1,25-(OH)2D3 receptors in other tissues. 相似文献
4.
Quantitation of vitamin D and its metabolites and their plasma concentrations in five species of animals 总被引:5,自引:0,他引:5
Chromatographic methods suitable for the resolution of 24,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, 25-hydroxyvitamin D3-26,23 lactone, and 25,26-dihydroxyvitamin D2 are described. These four metabolites comigrated in high-pressure liquid chromatography on silicic acid columns developed in 11:89 isopropanol:hexane. Adequate resolution was achieved by subjecting the four-metabolite complex to high-pressure liquid chromatography column developed in 2:98 isopropanol:methylene chloride. This additional chromatographic step, coupled with modifications of assay procedures previously described, allowed for the estimation of plasma concentrations of vitamin D2, vitamin D3, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D2, 24,25-dihydroxyvitamin D3, 25,26 dihydroxyvitamin D2, 25,26-dihydroxyvitamin D3, 25-hydroxyvitamin D3-26,23 lactone, and 1,25-dihydroxyvitamin D (1,25-dihydroxyvitamin D2 plus 1,25-dihydroxyvitamin D3). The samples automatically were introduced onto the high-pressure liquid chromatography columns with a Waters 710A “intelligent” processor. The metabolites were automatically collected with the aid of a programmable timer that advanced a fraction collector at predetermined intervals. The assays were used to determine the plasma vitamin D and vitamin D metabolite concentrations in five species of adult farm animals. 相似文献
5.
T A Reinhardt J L Napoli B Praminik E T Littledike D C Beitz J J Partridge M R Uskokovi? R L Horst 《Biochemistry》1981,20(21):6230-6235
A new metabolite of vitamin D3 has been isolated from the plasma of vitamin D3 treated cows and has been generated from 25(S),26-dihydroxyvitamin D3 with homogenates of vitamin D deficient chick kidney. This metabolite has been identified as 1,25,26-trihydroxyvitamin D3 by comigration with synthetic 1,25(S),26-trihydroxyvitamin D3 in four chromatographic systems, ultraviolet spectroscopy, mass spectrometry, and high-pressure liquid chromatography and mass spectrometry of derivatives. 1,25(S),26-Trihydroxyvitamin D3 is one-tenth as effective as 1,25-dihydroxyvitamin D3 in binding to the chick intestinal cytosol 1,25-dihydroxyvitamin D receptor. Either 25(S),26-dihydroxyvitamin D3 or 1,25-dihydroxyvitamin D3 can serve as precursor for in vitro production of 1,25,26-trihydroxyvitamin D3 by chick kidney tissue. 相似文献
6.
Out of Africa and back again: nested cladistic analysis of human Y chromosome variation 总被引:18,自引:3,他引:15
Hammer MF; Karafet T; Rasanayagam A; Wood ET; Altheide TK; Jenkins T; Griffiths RC; Templeton AR; Zegura SL 《Molecular biology and evolution》1998,15(4):427-441
We surveyed nine diallelic polymorphic sites on the Y chromosomes of 1,544
individuals from Africa, Asia, Europe, Oceania, and the New World.
Phylogenetic analyses of these nine sites resulted in a tree for 10
distinct Y haplotypes with a coalescence time of approximately 150,000
years. The 10 haplotypes were unevenly distributed among human populations:
5 were restricted to a particular continent, 2 were shared between Africa
and Europe, 1 was present only in the Old World, and 2 were found in all
geographic regions surveyed. The ancestral haplotype was limited to African
populations. Random permutation procedures revealed statistically
significant patterns of geographical structuring of this paternal genetic
variation. The results of a nested cladistic analysis indicated that these
geographical associations arose through a combination of processes,
including restricted, recurrent gene flow (isolation by distance) and range
expansions. We inferred that one of the oldest events in the nested
cladistic analysis was a range expansion out of Africa which resulted in
the complete replacement of Y chromosomes throughout the Old World, a
finding consistent with many versions of the Out of Africa Replacement
Model. A second and more recent range expansion brought Asian Y chromosomes
back to Africa without replacing the indigenous African male gene pool.
Thus, the previously observed high levels of Y chromosomal genetic
diversity in Africa may be due in part to bidirectional population
movements. Finally, a comparison of our results with those from nested
cladistic analyses of human mtDNA and beta-globin data revealed different
patterns of inferences for males and females concerning the relative roles
of population history (range expansions) and population structure
(recurrent gene flow), thereby adding a new sex-specific component to
models of human evolution.
相似文献
7.
JACQUES Destombes ET JEAN-LOUIS Henry 《Lethaia: An International Journal of Palaeontology and Stratigraphy》1987,20(2):129-139
A re-examination of the trilobite Baniaspis globosa Destombes (Phacopina) from the Ashgill of the Anti-Atlas (Morocco) shows that it has several derived characters which allow us to classify it in the family Calmoniidae. Calmoniids were characteristic members of the Malvino-Kaffric province during the Lower Devonian. Hence we regard the origins of this palaeogeographic province as located on the northern Gondwanan margin as early as Ashgill, or even Caradoc, times. We agree with those authors who have suggested that the trilobite associations present in the Anti-Atlas, the Montagne Noire (France) and Bohemia during the earlier Ordovician argue against the existence of a 'proto-Tethys' ocean. 相似文献
8.
JEAN-LOUIS HENRY ET JACQUES DESTOMBES 《Lethaia: An International Journal of Palaeontology and Stratigraphy》1991,24(3):249-253.
During the Llandeilo, the sandy lithological units which develop locally in the Armorican Massif, Spain and Morocco, are all characterized by similar trilobite associations in which endobenthic homalonotids. well adapted to shallow water and sandy substrate, are predominant. During the Llandeilo, on the northern Gondwanan margin, the wide geographical distribution of most of the trilobites, controlled by environmental factors, shows that the existence of a 'Proto-Tethys' ocean is unlikely. □ North Gondwana, Trilobita, Homalonotidae, Ordovician, Llandeilo. palaeoenvironment. 相似文献
9.
Herwig Bachmann Douwe Molenaar Michiel Kleerebezem Johan ET van Hylckama Vlieg 《The ISME journal》2011,5(5):929-932
Cooperative behavior is widely spread in microbial populations. An example is the expression of an extracellular protease by the lactic acid bacterium Lactococcus lactis, which degrades milk proteins into free utilizable peptides that are essential to allow growth to high cell densities in milk. Cheating, protease-negative strains can invade the population and drive the protease-positive strain to extinction. By using multiple experimental approaches, as well as modeling population dynamics, we demonstrate that the persistence of the proteolytic trait is determined by the fraction of the generated peptides that can be captured by the cell before diffusing away from it. The mechanism described is likely to be relevant for the evolutionary stability of many extracellular substrate-degrading enzymes. 相似文献
10.