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71.
Mesenchymal-epithelial conversions induced by 5-Azacytidine: Appearance of cytokeratin Endo-A messenger RNA 总被引:2,自引:0,他引:2
Alix Sémat Philippe Duprey Marc Vasseur Michel Darmon 《Differentiation; research in biological diversity》1986,31(1):61-66
When mouse-teratocarcinoma-derived fibroblasts (1246 cell line) are subjected to treatment with the inhibitor of DNA methylation, 5-Azacytidine (5 AzaC), they transiently express at 55-kilodalton intermediate-filament protein recognized by the epithelial-specific monoclonal antibody, TROMA-1, although they retain a fibroblastic morphology. However, rare clones (e.g., the 1339 cell line) that permanently express the antigen recognized by TROMA-1 can be derived from the 5 AzaC-treated 1246 population, and these clones have an epithelial phenotype. In the present study, we used cloned DNA probes to demonstrate that, in 1246 fibroblasts, 5 AzaC induces the appearance of Endo-A mRNA. High levels of Endo-A mRNA were also detected in the epithelial derivative, cell line 1339. In both cases, the capping site of the Endo-A mRNA was found to be the same as that in epithelial cells which normally express this RNA. 相似文献
72.
73.
Kinetics of malt starch hydrolysis by endogeneous alpha- and beta-amylases has been experimentally investigated in laboratory-, pilot- and industrial-scale reactors. The production rates of glucose, maltose, maltotriose and total extract, and the separate alpha- and beta-amylases deactivation rates are measured at varying mashing temperature and different initial starch concentrations and qualities. Based on the experimental results, a model is proposed that takes into account the initial carbohydrates and enzymes dissolution, the starch gelatinization, the separate hydrolytic action of alpha-and beta-amylases on insoluble and soluble starch and dextrins, and the influence of temperature both on enzyme activities and thermal denaturation rate. The model can predict, at the three scales, the final sugars concentrations in the wort for given initial malt concentrations and enzymatic contents, and for a fixed temperature profile during the mashing process. 相似文献
74.
To obtain a satisfactory agreement between computed transition temperatures and those determined experimentally, we introduce explicitly water molecules which hydrate the polar headgroup of dipalmitoylphosphatidylethanolamine molecules. The calculated free energy curves as a function of the intermolecular interchain distance and the degree of hydration of the polar groups permit the determination of the transition of the phospholipid system from the gel to the liquid crystalline phase. The detailed structure of the hydration shell is defined using the supermolecular approach. 相似文献
75.
We previously demonstrated inhibition of Na+-dependent 32Pi transport in canine renal brush-border membranes in association with NAD+-induced ADP ribosylation of membrane protein(s) and postulated that NAD+ inhibits Pi transport across the brush-border membrane via ADP ribosylation. Recently it was shown that incubation of rat brush-border membrane with NAD+ resulted in release of Pi which was prevented by EDTA. It was proposed that NAD+-mediated inhibition of 32Pi transport might occur through this mechanism. To determine whether NAD+ inhibited 32Pi transport by a mechanism other than or in addition to release of Pi, we compared Na+-dependent 32Pi counterflow in brush-border membrane equilibrated with Pi or with Pi generated from NAD+. Release of Pi from NAD+ incubated with brush-border membrane was confirmed. The increased uptake of 32Pi which was demonstrated in brush-border membrane equilibrated with Pi was not measured when intravesicular Pi was generated from a concentration of NAD+ which effected ADP-ribosylation of brush border membranes (100 μM NAD+). In contrast, increased uptake of 32Pi was demonstrated when intravesicular Pi was generated from 1 μM NAD+ which did not effect ADP ribosylation. Mg2+-dependent ADP ribosylation of brush-border membrane incubated with NAD+ was demonstrated which persisted during the time interval of 32Pi uptake measurements. Our findings are compatible with the hypothesis that NAD+-induced ADP ribosylation of brush-border membrane protein(s) results in inhibition of Pi transport across the membrane in vivo. EDTA may act to prevent this inhibition in brush-border membrane by chelation of Mg2+ and decreased ADP ribosylation. 相似文献
76.
Mavis Hendson Lizzie Askjaer Jennifer A. Thomson Marc van Montagu 《Applied microbiology》1983,45(5):1526-1532
Eighteen strains of Agrobacterium tumefaciens isolated from crown galls were tested for agrocin production. Of six agrocin-producing strains, one (D286) produced a broad-host-range agrocin active against strains carrying nopaline, octopine, and agropine type Ti plasmids. Sensitivity to agrocin D286 was found to map in the 11- to 18-megadalton region of the nopaline Ti plasmid pTiC58. The agrocin was partially purified, and its physical characteristics were consistent with its being a nucleotide, as is agrocin 84. Agrocin D286 was shown to inhibit DNA, RNA, and protein syntheses. Strain D286 spontaneously lost its pathogenicity, and its potential for use in the biological control of crown gall is discussed. 相似文献
77.
Genome structure of HBI, a variant of acute leukemia virus MC29 with unique oncogenic properties. 总被引:5,自引:3,他引:2
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We have analyzed the viral RNA of a variant of avian acute leukemia virus MC29, termed HBI. This virus was isolated during in vitro passage of a partially transformation-defective (td) mutant of MC29 (td10H-MC29) in chicken macrophages. While td10H-MC29 has a reduced ability to transform macrophages in vitro or to induce tumors in vivo, HBI-MC29 transforms macrophages efficiently and induces in vivo a high incidence of lymphoid tumors. Electrophoretic analysis of HBI-MC29 genomic RNA revealed that it has a complexity of 5.7 kilobases, like the RNA of wild-type (wt) MC29, and that it is 0.6 kilobases longer than the 5.1-kilobase RNA of the deletion mutant td10H-MC29. Analysis of the viral RNAs of two clonal isolates of HBI-MC29 by T1 oligonucleotide fingerprinting showed that sequences from the viral transformation-specific region, v-myc, which are deleted in td10H RNA, are present in HBI RNA. Moreover, hybridization of HBI RNA to molecularly cloned subgenomic fragments of wtMC29 proviral DNA, followed by fingerprint analysis of hybridized RNA, showed that the entire v-myc-specific RNA sequences defined previously are present. Hybridization to cloned DNA of the normal chicken locus c-myc shows a close relationship between HBI v-myc RNA and c-myc DNA, especially in the sequences which were deleted from td10H-MC29. T1 oligonucleotide maps of HBI and td10H RNAs were prepared and compared. Total conservation of the oligonucleotide pattern is observed in the overlapping v-myc regions, while the partial structural genes gag and env show some variations, most of which can be directly proven to be due to point mutations or recombination with helper viral RNAs that were analyzed in parallel. Recombination of td10H-MC29 with c-myc, followed by recombinational and mutational changes in the structural genes during passage with helper virus, could be a possible explanation for the origin of HBI. 相似文献
78.
A simple procedure for the isolation and preparative gel electrophoresis of snRNA and scRNA is described. These small RNA species were used to select DNA sequences from a human genomic library which are able to protect hybridized snRNA or scRNA against T1-ribonuclease attack. The snRNA clones obtained contain only sequences for one snRNA species and only one copy of the respective gene. In contrast, more than one 7S RNA gene is present within the scRNA clones. 相似文献
79.
The genes coding for histone H3 and H4 in Neurospora crassa are unique and contain intervening sequences. 总被引:39,自引:14,他引:25
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L P Woudt A Pastink A E Kempers-Veenstra A E Jansen W H Mager R J Planta 《Nucleic acids research》1983,11(16):5347-5360
Sequences coding for histone H3 and H4 of Neurospora crassa could be identified in genomic digests with the use of the corresponding genes from sea urchin and X. laevis as hybridization probes. A 2.6 kb HindIII-generated N. crassa DNA fragment, showing homology with the heterologous histone H3-gene probes was cloned in a charon 21A vector. Using DNA from this clone as a homologous hybridization probe a 6.9 kb SalI-generated DNA fragment was isolated which in addition to the histone H3-gene also contains the gene coding for histone H4. Several lines of evidence demonstrate the presence of only a single histone H3- as well as a single histone H4-gene in N. crassa. The two genes are physically linked on the genome. DNA sequencing of the N. crassa histone H3- and H4-genes confirmed their identity and, in addition, revealed the presence of one short intron (67 bp) within the coding sequence of the H3-gene and even two introns (68 and 69 bp) within the H4-gene. The amino acid sequences of the N. crassa histones H3 and H4, as deduced from the DNA sequences, and those of the corresponding yeast histones differ only at a few positions. Much larger sequence differences, however, are observed at the DNA level, reflecting a diverging codon usage in the two lower eukaryotes. 相似文献
80.