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排序方式: 共有88条查询结果,搜索用时 15 毫秒
1.
D. Broekaert K. Cooreman P. Coucke S. Nsabumukunzi P. Reyniers P. Kluyskens E. Gillis 《The Histochemical journal》1982,14(4):573-584
Summary A quantitative histochemical study was carried out on the distribution of protein thiol and disulphide groups in normal human plantar epidermal tissue. Histochemical demonstration of reactive groups was achieved by addition ofN-(4-aminophenyl) maleimide, subsequent diazotization and final coupling with a Nitro Red or chromotropic acid label as first described by Sippel. The quantitative reliability of the method was tested by absorption cytophotometry, and evaluated on the basis of the internal consistency of the results reported.Our histological observations and histophotometric data support accepted views on epidermal keratinization. A limited, though reproducible, amount of disulphide bonds was observed near the basement membrane. The free thiol concentration in basal and prickle cells was low and almost constant, but was higher in the granular cells, where deposition of sulphur-containing proteins on cell membranes is initiated. In Malpighian layers, disulphide cross-links only occurred just beneath the transition zone in thickened cell membranes. The staining pattern of the inner stratum corneum resembled a mosaic and was characterized by a sharp rise of the disulphide content, which exceeded the decrease in free thiol groups. The free thiol concentration decreased further throughout the cornified layers whilst the disulphide content remained fairly constant. Staining of thiol and disulphide groups together corresponded, within the limits of the standard error, to the sum of the thiol and disulphide concentrations when they were assayed separately in living and horny cells. These results confirm that living cells are the main site of free thiol groups, while horny cells are the most prominent site of disulphide cross-links. 相似文献
2.
3.
Gianmarco Rinaldi Erica Pranzini Joke Van Elsen Dorien Broekaert Cornelius M. Funk Mélanie Planque Ginevra Doglioni Patricia Altea-Manzano Matteo Rossi Vincent Geldhof Shao Thing Teoh Christina Ross Kent W. Hunter Sophia Y. Lunt Thomas G.P. Grünewald Sarah-Maria Fendt 《Molecular cell》2021,81(2):386-397.e7
4.
Grebe M Pröfrock D Kakuschke A Broekaert JA Prange A 《Metallomics : integrated biometal science》2010,2(10):683-693
The health status of marine mammals such as harbour seals (Phoca vitulina) represents an indirect but powerful way for the assessment of environmental changes. The present work illustrates the first investigation and characterisation of Tf isolated from blood samples of North Sea harbour seals with a view to using changes in Tf isoform patterns as an additional parameter in extended studies of their health status. Therefore, an HPLC-ICP-MS approach has been developed which allows the highly resolved separation and fractionation of up to eight different Tf isoforms, as well as their sensitive and specific detection on the basis of their characteristic iron content. Molecule-specific detection techniques such as nanoLC-ESI-QTRAP-MS or MALDI-TOF-MS were used as complementary techniques to unambiguously identify the isolated proteins as Tf via cross species protein identification and to further characterise the molecular weight as well as the sialic acid content, which is responsible for the elution behaviour of the different isoforms during their ion exchange separation. A molecular mass above 80 kDa has been measured for the different seal Tf isoforms, which is in good agreement with the known molecular mass in other mammalian species, while the estimated pI of the different isoforms indicates some differences in comparison to other species. A number of homologies to known Tf sequences have been observed, which finally allows the cross species protein identification. The combined metallomics orientated analytical approach, which includes the complementary application of element and molecule-specific detection techniques, opens up interesting possibilities for the fast and targeted isolation and identification of a diagnostically relevant metal containing protein from an un-sequenced mammalian species prior to its utilisation in extended studies. 相似文献
5.
Tierens KF Thomma BP Bari RP Garmier M Eggermont K Brouwer M Penninckx IA Broekaert WF Cammue BP 《The Plant journal : for cell and molecular biology》2002,29(2):131-140
An Arabidopsis thaliana mutant, esa1, that shows enhanced susceptibility to the necrotrophic pathogens Alternaria brassicicola, Botrytis cinerea and Plectosphaerella cucumerina, but has wild-type levels of resistance to the biotrophic pathogens Pseudomonas syringae pv. tomato and Peronospora parasitica. The enhanced susceptibility towards necrotrophic pathogens correlated with a delayed induction of phytoalexin accumulation and delayed induction of the plant defensin gene PDF1.2 upon inoculation with pathogens. Two reactive oxygen generating compounds, paraquat and acifluorfen, were found to cause induction of both phytoalexin accumulation and PDF1.2 expression in wild-type plants, but this induction was almost completely abolished in esa1. This finding suggests that esa1 may somehow be involved in transduction of signals generated by reactive oxygen species. 相似文献
6.
Pathogen-induced systemic activation of a plant defensin gene in Arabidopsis follows a salicylic acid-independent pathway. 总被引:21,自引:0,他引:21 下载免费PDF全文
I A Penninckx K Eggermont F R Terras B P Thomma G W De Samblanx A Buchala J P Mtraux J M Manners W F Broekaert 《The Plant cell》1996,8(12):2309-2323
A 5-kD plant defensin was purified from Arabidopsis leaves challenged with the fungus Alternaria brassicicola and shown to possess antifungal properties in vitro. The corresponding plant defensin gene was induced after treatment of leaves with methyl jasmonate or ethylene but not with salicylic acid or 2,6-dichloroisonicotinic acid. When challenged with A. brassicicola, the levels of the plant defensin protein and mRNA rose both in inoculated leaves and in nontreated leaves of inoculated plants (systemic leaves). These events coincided with an increase in the endogenous jasmonic acid content of both types of leaves. Systemic pathogen-induced expression of the plant defensin gene was unaffected in Arabidopsis transformants (nahG) or mutants (npr1 and cpr1) affected in the salicylic acid response but was strongly reduced in the Arabidopsis mutants eln2 and col1 that are blocked in their response to ethylene and methyl jasmonate, respectively. Our results indicate that systemic pathogen-induced expression of the plant defensin gene in Arabidopsis is independent of salicylic acid but requires components of the ethylene and jasmonic acid response. 相似文献
7.
An De Bondt Kristel Eggermont Iris Penninckx Inge Goderis Willem F. Broekaert 《Plant cell reports》1996,15(7):549-554
We have previously developed a protocol for efficient gene transfer and regeneration of transgenic calli following cocultivation of apple (cv. Jonagold) explants with Agrobacterium tumefaciens (De Bondt et al. 1994, Plant Cell Reports 13: 587–593). Now we report on the optimization of postcultivation conditions for efficient and reproducible regeneration of transgenic shoots from the apple cultivar Jonagold. Factors which were found to be essential for efficient shoot regeneration were the use of gelrite as a gelling agent and the use of the cytokinin-mimicing thidiazuron in the selective postcultivation medium. Improved transformation efficiencies were obtained by combining the hormones thidiazuron and zeatin and by using leaf explants from in vitro grown shoots not older than 4 weeks after multiplication. Attempts to use phosphinothricin acetyl transferase as a selectable marker were not successful. Using selection on kanamycin under optimal postcultivation conditions, about 2% of the leaf explants developed transgenic shoots or shoot clusters. The presence and expression of the transferred genes was verified by -glucuronidase assays and Southern analysis. The transformation procedure has also been succesfully applied to several other apple cultivars.Abbreviations BAP
benzylaminopurine
- CTAB
hexadecyltrimethylammoniumbromide
- Na2EDTA
ethylenediamine-tetra-acetate ferric-sodium salt
- FeNaEDTA
ethylenediamine-tetra-acetate ferric-sodium salt
- GA3
gibberellic acid 3
- GusA
-glucuronidase
-
gusA
-glucuronidase gene of Escherichia coli
- IAA
indole acetic acid
- IBA
indole butyric acid
- 2iP
N6-2-isopentenyl adenine
- NAA
naphthalene acetic acid
-
nptII
neomycinphosphotransferase II gene
-
bar
phosphinothricin acetyl transferase gene
- PCR
polymerase chain reaction
- PPT
phosphinothricin
- STS
silver thiosulphate
- T-DNA
transferred DNA
- TDZ
thidiazuron
- X-Gluc
5-bromo-4-chloro-3-indolyl -D-glucuronide
- Zea
trans-Zeatin 相似文献
8.
Kristel Eggermont Inge J. Goderis Willem F. Broekaert 《Plant Molecular Biology Reporter》1996,14(3):273-279
We describe here a reliable high-throughput method for extraction of RNA from fresh or frozen plant tissue that obviates laborious
and time-consuming homogenisation by mortar and pestle. The method is based on homogenisation by high-speed reciprocal shaking
in presence of a mixture of inexpensive abrasive materials; i.e., quartz sand and glass beads. After homogenisation, the method
follows a standard procedure for RNA extraction by phenol/LiCl. Yield and quality of RNA obtained by homogenisation with the
sand/glass bead mix are identical to those obtained by mortar and pestle. 相似文献
9.
A lectin has been isolated from rhizomes of ground elder (Aegopodium podagraria) using a combination of affinity chromatography on erythrocyte membrane proteins immobilized on cross-linked agarose and hydroxyapatite, and ion-exchange chromatography. The molecular structure of the lectin was determined by gelfiltration, sucrose density-gradient centrifugation and gel electrophoresis under denaturing conditions. It has an unusually high Mr (about 480000) and is most probably an octamer composed of two distinct types of subunits with slightly different Mr (about 60000). Hapten inhibition assays indicated that the Aegopodium lectin is preferentially inhibited by N-acetylgalactosamine. Nevertheless, it does not agglutinate preferentially blood-group-A erythrocytes. The ground-elder lectin is a typical non-seed lectin, which occurs virtually exclusively in the underground rhizomes. In this organ it is an abundant protein as it represents up to 5% of the total protein content. The lectin content of the rhizome tissue varies strongly according to its particular location along the organ. In addition, the lectin content changes dramatically as a function of the seasons. The ground-elder lectin differs from all other plant lectins by its unusually high molecular weight. In addition, it is the first lectin to be isolated from a species of the family Apiaceae.Abbreviations APA
Aegopodium podagraria agglutinin
- PBS
phosphate-buffered saline
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis 相似文献
10.