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61.
Summary We have introduced the gene encoding luciferase from Photinus pyralis into pear and tobacco cells in order to judge the reaction of plant tissue to damaging conditions such as incubation at high temperature or inoculation with a pathogen. The constitutive expression of the luciferase gene via a strong promoter slowly decreased during propagation of the transformed pear cell line. After various stress treatments the resulting luciferase activity and the ATP content of the plant cells were determined by bioluminescence and found to correspond to each other. Inoculation of transformed pear cells with Erwinia amylovora resulted in a continuous decrease of luciferase activity in contrast to tobacco cells, where the enzyme activity was significantly higher in the first period after inoculation with bacteria compared to the untreated control cells. The pattern of the luciferase activity reflected the slow damage of the host-plant cells by E. amylovora and the elevated metabolism of the non-host cells after inoculation with the pathogen.Abbreviations 2,4-D
2,4-dichloro-phenoxyacetic acid
- CaMV
Cauliflower mosaic virus
- DTT
dithiothreitol
- EDTA
ethylenediaminetetraacetate
- FDA
fluorescein diacetate
- HEPES
(hydroxyethyl)piperazine(ethanesulfonic acid)
- HR
hypersensitive reaction
- Tris
tris (hydroxymethyl)amino-methane 相似文献
62.
63.
Izaura Yoshico Hirata Maria Helena Sedenho Cezari Clovis Ryuichi Nakaie Paulo Boschcov Amando Siuiti Ito Maria Aparecida Juliano Luiz Juliano 《Letters in Peptide Science》1995,1(6):299-308
Summary A general procedure, using the commonly employed solid-phase peptide synthesis methodology for obtaining internally quenched fluorogenic peptides with ortho-aminobenzoyl/dinitrophenyl groups as donor-acceptor pairs, is presented. The essential feature of this procedure is the synthesis of an N
-Boc or-Fmoc derivative of glutamic acid with the -carboxyl group bound to N-(2,4-dinitrophenyl)-ethylenediamine (EDDnp), which provides the quencher moiety attached to the C-terminus of the substrate. The fluorescent donor group, ortho-aminobenzoic acid (Abz), is incorporated into the resin-bound peptide in the last coupling cycle. Depending on the resin type used, Abz-peptidyl-Gln-EDDnp or Abz-peptidyl-Glu-EDDnp is obtained. Using the procedure described above, substrates for human renin and tissue kallikreins were synthesised. Spectrofluorimetric measurements of Abz bound to the -amino group of proline showed that strong quenching of Abz fluorescence occurs in the absence of any acceptor group. 相似文献
64.
Cloning and mapping of Saccharomyces cerevisiae photoreactivation gene PHR1. 总被引:9,自引:3,他引:6 下载免费PDF全文
The yeast Saccharomyces cerevisiae, like most organisms, is able to directly repair pyrimidine dimers by using a photoreactivating enzyme and visible light. Cells carrying the phr1 mutation were shown previously to be unable to photoreactivate dimers, but neither the map position nor the primary gene product of the PHR1 gene has been determined. We have cloned this gene and determined its map position. A plasmid containing a 6.4-kilobase yeast DNA insert has been isolated and shown to restore photoreactivation in a phr1 strain. A 3.1-kilobase subclone has also been shown to complement phr1. The original plasmid was targeted to integrate into chromosomal DNA at a site homologous to the insert by cutting within the insert. Two of these integrants have been mapped on the right arm of chromosome XV; the integrants have been further mapped at ca. 13 centimorgans from prt1. It has also been independently determined that phr1 maps at this location. Thus, we have determined the map position of PHR1 and also have shown that the plasmid contains PHR1 rather than a suppressor of the phr1 mutation. 相似文献
65.
66.
William r. Anderson Willi Frick G.Doyle Daves Douglas F. Barofsky Isomaro Yamaguchi Ding Chang Karl Folkers Sune Rosell 《Biochemical and biophysical research communications》1977,78(1):372-376
Mass spectra of underivatized hexa- and heptapeptide amides related to Substance P have been obtained with a conventional electron ionization mass spectrometer using sample vaporization from a tungsten wire by the technique of rapid heating, proton transfer ionization using ammonia, and photoplate recording of spectra. These spectra exhibit little evidence of sample pyrolysis and are readily interpreted to yield amino acid sequences. 相似文献
67.
The association of [3H] [D-Ala2, D-Leu5] enkephalin ([3H]DADLE]) with mouse neuroblastoma cells (N4TG1) was investigated. Under identical conditions the time course, dose response curve and temperature dependence for ligand uptake were similar to those for ligand-induced receptor loss (down regulation). Uptake of [3H]DADLE was inhibited by opiate ligands as well as by the metabolic inhibitors sodium azide and 2,4 dinitrophenol. Comparison of the effects of these inhibitors on receptor binding, ligand uptake and receptor loss indicated that these cells accumulate [3H]DADLE in excess of their surface receptor number. The data suggest that receptor recycling occurs and that ligand is internalized via receptor mediated endocytosis. 相似文献
68.
69.
Chang CW 《Plant physiology》1979,63(5):973-977
During cotton leaf development, starch accumulation was characterized by an initial rise to a maximum at the second to the fourth leaf from the apex. Then, starch content progressively decreased with leaf age. Starch accumulation was inversely related to the ratio of amylopectin to amylose. Differences between leaves in this ratio resulted from variations in both amylose and amylopectin levels. Fluctuations in amylose levels were more extreme than those of amylopectin. 相似文献
70.