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1.
PCR products were characterized by electrophoresis, blotting and hybridization. In addition to the bands of expected size, bands of slower electrophoretic mobility were often detected. The slower bands completely disappeared when the PCR products were subjected to slow cooling, treated with S1 nuclease or run on an alkaline gel, whereas the bands of expected size were unaffected. The slower bands are therefore likely to contain single-stranded DNA. 相似文献
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3.
Marion H. Brown Patricia A. Gorman William A. Sewell Nigel K. Spurr Denise Sheer Michael J. Crumpton 《Human genetics》1987,76(2):191-195
Summary A cDNA clone encoding the human T lymphocyte sheep erythrocyte receptor [the CD2 (T11) antigen] was used as a probe to define the chromosomal location of the gene. The signal, revealed by hybridisation to Southern blots of genomic DNA from somatic cell hybrids, showed a high degree of concordance for human chromosome 1. In particular, the hybrid F4Sc13C19 which contained the short arm only of human chromosome 1 was positive. The location of the CD2 gene to 1p13 was confirmed by in situ hybridisation. 相似文献
4.
A hybrid histone octamer was reconstituted from erythrocyte H2A and H2B, avian [110 Cys-des-thio]histone H3 and the sea-urchin sperm [73Cys]H4 variant. [110Cys-Des-thio]histone H3 was prepared by reaction of natural H3 with Raney nickel. The ability of the hybrid octamer to crystallize to the same form as the natural octamer demonstrated that the chemical modification of cysteine to alanine in H3 and the mutation from threonine to cysteine in sperm H4 do not alter histone-histone interactions in the octamer. Since the sulfhydryl groups of both H4 molecules are fully accessible to 5,5'-dithiobis(2-nitrobenzoate) these residues provide suitable sites for the introduction of a single cysteine-specific label per H4 molecule in the octamer. 相似文献
5.
D Landsman B T Sewell C von Holt 《Biochemical and biophysical research communications》1988,155(1):66-73
The pancreatic deoxyribonuclease (DNase I) digestion rates at the susceptible sites on nucleosomal core particles from blastula, gastrula and sperm cells of the sea urchin, Parechinus angulosus, have been determined. Although there are differences in their isohistone composition, the rates of digestion are similar for both embryonic stages. The rates of digestion for sperm core particles are 3-5 times lower than for embryo core particles at the more, and up to 2.5 times lower at the less susceptible sites. An explanation for these differences could be sought in the sperm isohistones H2B which are characterized by N-terminal extensions of 20-25 amino acid residues. 相似文献
6.
We have studied the effects on interferon-gamma (IFN-gamma) production of pertussigen, a protein toxin from Bordetella pertussis that augments and prolongs delayed-type hypersensitivity (DTH) reactions. Lymphoid cell suspensions from immunized mice were incubated with antigen or mitogen, and the culture supernatants were assayed for IFN-gamma. The production of IFN-gamma on exposure to specific antigen or concanavalin A was greatly enhanced if mice were given pertussigen at the time of immunization. There was no detectable IFN-gamma production when cells were exposed to saline or to an irrelevant antigen. The effect of pertussigen on antigen-driven IFN-gamma production correlated with its effect on the capacity of the same cell populations to transfer DTH. The enhanced IFN-gamma production by cells from mice given pertussigen could not be attributed to an increased antigen-presenting capacity of this cell population. Production of IFN-gamma was abolished if the cells were pretreated with emetine, but not with mitomycin C, and the release of IFN-gamma was not detected in the first 8 hr of culture. After immunization with pertussigen, IFN-gamma was produced by lymph node and spleen cells from 7 days onward and both cell types produced IFN-gamma until at least 30 days after immunization. It is suggested that the augmentation of antigen-specific IFN-gamma production may contribute to the prolonged DTH reactions induced by pertussigen in vivo. 相似文献
7.
H J Greyling J P Hapgood B T Sewell C von Holt 《European journal of biochemistry》1986,161(1):133-138
Histone octamers were covalently labelled with aurothiomalate at amino groups by the method of carbodiimide activation. The labelling procedure was demonstrated to result in the specific covalent coupling through a single bond of the heavy metal atom label to protein amino groups. Such octamers were dissociated to yield soluble H2A-H2B dimers containing three gold atoms per dimer. The dimers were reconstituted with native H3-H4 tetramers to form labelled octamers, which were crystallized to form helical tubes. This strongly suggests that this procedure resulted in minimal changes of protein conformation. 相似文献
8.
Rehydration and equilibration (4 h) of feeds and feed ingredients at water/sample ratios (vol/wt) of 1.4 to 3.2 did not markedly increase recovery of Salmonella spp. in the slurry when analyzed by standard cultural and direct enrichment methods. Of 143 naturally contaminated samples examined, equilibration increased levels of detection from 106 to 109 positive samples by the standard cultural method and from 103 to 112 by direct enrichment. Results suggest that nonhomogeneous distribution of low incident numbers of salmonellae in test samples rather than an equilibration-dependent response provided for the observed heterogeneity in recovery patterns. The novel equilibration approach is of limited application and requires validation on an individual food basis. 相似文献
9.
Rehydration and equilibration (4 h) of feeds and feed ingredients at water/sample ratios (vol/wt) of 1.4 to 3.2 did not markedly increase recovery of Salmonella spp. in the slurry when analyzed by standard cultural and direct enrichment methods. Of 143 naturally contaminated samples examined, equilibration increased levels of detection from 106 to 109 positive samples by the standard cultural method and from 103 to 112 by direct enrichment. Results suggest that nonhomogeneous distribution of low incident numbers of salmonellae in test samples rather than an equilibration-dependent response provided for the observed heterogeneity in recovery patterns. The novel equilibration approach is of limited application and requires validation on an individual food basis. 相似文献
10.
Z Yu P S Williams H J Smith R D Sewell A Patel J P Gonzalez 《Journal of enzyme inhibition》1989,3(2):103-117
Some 2-substituted-(2'-aminophenyl)-4-thioxohydantoic acids (o-amino PTC-amino acids) have antinociceptive activity when administered (icv) alone (IC50 = 0.04-0.87 microM/animal) and show a striking prolongation of the antinociceptive action of (D-Ala-2 D-Leu5)-enkephalin (DADL) in combination. The effects are thought to be mediated via opioid receptors since they are naloxone-reversible. Although inhibitors of the enkephalin degrading puromycin-insensitive, bestatin-sensitive aminopeptidase (possibly aminopeptidase M) their action is weak (IC50 = 32 microM leucine, 536 microM, glycine) and they might be considered to have a direct antinociceptive effect on opioid receptors. The titled compounds constitute novel 'lead' compounds for the development of potent aminopeptidase M inhibitors. 相似文献