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1.
Dr Jean-Charles Cailliez Cristina Cantelli Nathalie Séguy Stefania Conti Mara Gerloni Giulia Morace Luciano Polonelli 《Mycopathologia》1994,126(3):173-177
A secreted killer toxin was detected through the cell wall ofPichia anomala cells by ultrastructural immunodetection with a specific monoclonal antibody (MAb KT4). MAb KT4 was successively detected by colloidal gold labeled streptavidin and biotinylated anti-mouse F(ab')2 antibodies. The antigenic determinants of the toxin were localized throughout the cytoplasm and the cell wall of killer yeast cells. The Lowicryl K4M-immunogold method gave very satisfactory results and showed that the killer toxin was somewhat concentrated in the yeast cell wall layers before being exported into the medium. In agreement with previous reports, the binding of MAb KT4 suggested that theP. anomala killer toxin secretion did not result from a homogeneous diffusion across the yeast cell wall.Abbreviations G15
gold particles of 15 nm
- IEM
immunoelectron microscopy
- IFA
immunofluorescence assay
- MAb
monoclonal antibody
- PBS
phosphate buffered saline
- SAM/F(ab)2
sheep antibodies anti-mouse F(ab)2
- SBB
Sabouraud buffered broth 相似文献
2.
Autosporulation is a common mode of propagation for unicellular algae. Autospore-forming species of Chlorellaceae, Chlorella vulgaris Beijerinck, C. sorokiniana Shihira et Krauss, C. lobophora Andreyeva, and Parachlorella kessleri (Fott et Nováková) Krienitz et al. have glucosamine as the main constituent of their rigid cell wall. Recent phylogenetic analyses have showed that the Chlorellaceae divided into two sister groups: the Chlorella-clade and the Parachlorella-clade. We compared the cell wall structure and synthesis of the daughter cell wall in the four species by electron microscopy using rapid freezing and freeze substitution methods. The cell wall of C. vulgaris, C. sorokiniana, and C. lobophora consisted of an electron-dense thin layer with an average thickness of 17–20, 22, and 19 nm, respectively. In these three species, daughter cell wall synthesis occurred on the outer surface of the plasma membrane in the early cell-growth phase. The cell wall of P. kessleri, however, was electron-transparent and 54–59 nm in thickness. Ruthenium red staining of P. kessleri indicated that ruthenium-red-specific polysaccharides accumulated over the outer surface of the plasma membrane. Immunoelectron microscopic observation with an anti--1, 3-glucan antibody and staining with wheat germ agglutinin (WGA) indicated that the cell wall contained -1, 3-glucan and WGA specific N-acetyl--D-glucosamine. In P. kessleri, daughter cell wall synthesis began after successive protoplast division. The daughter cell wall synthesis during autosporulation in the four species of Chlorellaceae can be classified into two types—the early and the late types. 相似文献
3.
Summary The maize root cap is a tissue known for its high production of a fucose-rich slime. At the cellular periphery, two kinds of components exist which are indistinguishable: the cell wall barrier and the slime which passes through. Two complementary probes were used, both at the light and the electron microscope level, in order to distinguish the different components. The lectinUlex europaeus agglutinin I was used as a probe targetting the slime and the enzyme cellulose 1,4--D-glucan cellobiohydrolase I was used to probe the cellulose framework. Both probes were used either alone or sequentially for double labeling. The cytochemical PATAg test was optionally used with the enzyme-gold complex labeling. After several technical improvements (multistep method, increase in accessibility), UeA I was used to follow the exocytic pathway of the slime from the Golgi apparatus to the exterior of the cell. The results indicate the occurrence of at least two populations of Golgi apparatus vesicles, and one is directly engaged in the transport of the fucoserich slime. The slime accumulated in pockets between the plasmamembrane and the outer tangential cell wall. The CBH I-gold complex showed the existence and the maintenance of a thin but continuous cellulosic layer, even when the cells slough. The double labeling showed the fucose-rich compounds within the cell wall. Data emphasize the role of the cell wall as a filtering barrier and a mechanical protection in the course of differentiation.Abbreviations CBH I
EC 3.2.1.91, cellulose 1,4--D-glucan cellobiohydrolase I
- CMC
carboxymethyl cellulose
- CPB
citrate phosphate buffer
- FITC
fluorescein isothiocyanate
- IC
internal cell
- PA-TAg
periodic acid-thiocarbohydrazide-silver proteinate
- PBS
phosphate buffered saline
- PC
cell with accumulation pockets
- PEG
polyethyleneglycol
- SC
sloughed cell
- UeA I
Ulex europaeus agglutinin I
- VI
UeA I-labelled Golgi-derived vesicles
- V2
UeA I-unlabelled Golgi-derived vesicles 相似文献
4.
Galactomannan was localized by scanning and transmission electron microscopy on the cells and cell walls of Schizosaccharomyces pombe. The markers were prepared from colloidal gold granules labelled with an -galactopyranosyl-binding lectin isolated from the seeds of Bandeiraea simplicifolia. Part or all of this -galactomannan was present in the outer layer of the cell wall and was uniformly distributed even on the fission scars.Non-Standard Abbreviations Au
lectin-labelled colloid
- SEM
scanning electron microscopy
- TEM
transmission electron microscopy 相似文献
5.
An attachment tip and pili-like structures in insect- and plant-pathogenic spiroplasmas of the class Mollicutes 总被引:1,自引:0,他引:1
Ultrastructural studies using scanning electron microscopy (SEM), negative-staining transmission electron microscopy (TEM), and thin-sectioning TEM on four species of Spiroplasma, in vitro and/or in vivo, indicated that their helices commonly possess one tapered end (tip structure) and one blunt or round end. These tip structures appeared morphologically different from the rest of the helix, exhibiting an electron-dense conical or rod-shaped core. In thin sections of the midgut of the leafhopper Dalbulus elimatus, the tip structures of Spiroplasma kunkelii in the midgut lumen were mostly aligned between microvilli, perpendicular to the apical plasma membrane of epithelial cells. These tip structures appeared frequently attached or closely apposed to the plasma membrane, in which cup-shaped invaginations close to the tips were observed. Pleomorphic forms of spiroplasma, enclosed in membranous vesicles, were found in the cytoplasm of the midgut epithelial cells. These findings suggest that the tip structure may be involved in the orientation and attachment of spiroplasma helices in relation to their host cells, and thus may be functionally comparable to the attachment organelle of mycoplasmas. Additionally, pili-like structures were observed by negative-staining TEM on the surface of Spiroplasma melliferum, and in thin sections of S. kunkelii infecting the leafhopper vector Dalbulus gelbus.
Abbreviations
CSS
Corn stunt spiroplasma
-
SEM
Scanning electron microscopy
-
TBS
Tris-buffered saline
-
TEM
Transmission electron microscopy 相似文献
6.
Summary The ultrastructure and composition of the extracellular matrices (ECMs) associated with germ tubes and appressoria ofColletotrichum lindemuthianum have been examined. Flexuous fibres (fimbriae), up to 6 m long and 4–30 nm in diameter, protruded from the surface of germ tubes and appressoria. Anionic colloidal gold and lectin cytochemistry showed that ECMs of germ tubes and appressoria contain basic proteins, -D-mannose and -D-galactose residues. A monoclonal antibody, UB26, was raised to infection structures isolated from leaves ofPhaseolus vulgaris infected withC. lindemuthianum. UB26 recognised a protein epitope on two glycoproteins (Mr 133,000 and 146,000). Reductions in the Mr of these proteins after treatment with peptide-N-glycosidase and trifluoromethane sulphonic acid suggest that they carry N- and O-linked side-chains. Immunofluorescence and EM-immunogold labelling showed that glycoproteins recognised by UB26 were restricted to the ECMs around germ tubes and appressoria but fimbriae were not labelled. Unlike appressorial germ tubes formed in vitro, intracellular infection hyphae were not labelled, suggesting that the glycoproteins recognised by UB26 are not present on fungal structures formed within host cells. In liquid culture, these glycoproteins were not released into the medium, suggesting they are physically linked to the cell wall. Also, the glycoproteins were not removed from glass surfaces by ultrasonication. These results suggest that glycoproteins recognised by UB26 may be involved in the adhesion of germ tubes and appressoria to substrata. Our results show that the ECMs of germ tubes and appressoria differ markedly in structure and composition from those of conidia and intracellular hyphae, and that extracellular glycoproteins are associated with specific regions of the fungal cell surface.Abbreviations ECM
extracellular matrix
- BPA
Bauhinia purpurea agglutinin
- BSA
bovine serum albumin
- DIC
differential interference contrast
- FITC
fluorescein isothiocyanate
- GNL
Galanthus nivalis lectin
- GSI-B4
Griffonia simplicifolia isolectin B4
- HEPES
(N-(2-hydroxyethyl)piperazine-N-(2-ethanesulphonic acid)
- IIF
indirect immunofluorescence
- IPC
isopycnic centrifugation
- MAb
monoclonal antibody
- PEG
polyethylene glycol
- PBS
phosphate buffered saline
- PNGase
peptideN-glycosidase
- SDS-PAGE
sodium dodecyl sulphate polyacrylamide gel electrophoresis
- TCS
tissue culture supernatant
- TEM
transmission electron microscopy
- TFMS
trifluoromethane sulphonic acid 相似文献
7.
Wheamei Jenq Chi Lan Chen Chun Chang Chang Richard F. E. Crang 《Archives of microbiology》1994,162(1-2):33-40
Selected strains of Candida albicans were examined to reveal the surface antigenicity and biochemical nature of major cell wall proteins that also were shown to serve as cellular adhesins on human buccal epithelial cells. Confirmation of the adhesive properties of these cells was made by scanning electron microscopy and immunofluorescence microscopy. Particular attention was directed at the clinical isolate KM-302. By means of indirect immunofluorescence staining, the KM-302 blastoconidia absorbed rabbit anti-C. albicans ATCC-32354 serum, revealing specific localization of surface antigens on germ tubes and pseudohyphae. Extracellular polymeric material and the cell wall extract of C. albicans KM-302 blastoconidia were found to contain a major surface antigen of 49 kDa that exhibited 42% adhesion inhibition in vitro. Of considerable significance is that immunogold localization by electron microscopy showed the antigen to be almost exclusively cell wall bound. This major antigen, identified in affinity and gel filtration chromatography fractions, was composed of 4% carbohydrate and 95.7% protein and had an isoelectric point of 6.1. The major antigen also showed a high level of similarity with that of C. albicans strain SC-5314 inasmuch as the major antigen of that strain had carbohydrate and protein compositions of 4 and 95.5%, respectively. Both of these strains also possessed the same percent of adhesion inhibition of human buccal epithelial cells.Abbreviations
BECs
buccal epithelial cells
-
CWE
cell wall extract
-
EPP
extracellular polymers and proteins
-
FITC
fluorescein isothiocyanate
-
mAg
major antigen
-
OD
600
optical density at 600 nm
-
PBS
phosphate buffered saline
-
TEM
transmission electron microscopy
-
YNB
yeast nitrogen base 相似文献
8.
Summary The endobiotic thallus ofPhysoderma maydis is characterized by the presence of an extremely fine rhizomycelium which passes through the host cell wall, allowing the spread of the disease, and irregularly shaped turbinate cells, which may be septate or nonseptate and which are in close association with developing resting sporangia. The formation of the resting sporangium wall is first seen as localized depositions on the rounded surface of the sporangium and only later on the flattened surface of the sporangium which will form the operculum. The substructure of the resting sporangium wall is typical for members of theBlastocladiales. The resting sporangium is contiguous with the rhizomycelium during development and is finally sealed-off from the rhizomycelium by a further deposition of wall material. After the sealing-off of the resting sporangium from the rhizomycelium the content of the sporangium is compartmentalized and the two inner wall layers are deposited. The centre of the sporangium is filled with an electron dense accretion. At the periphery of the sporangium is a layer of lipid bodies. Between the lipid bodies and the central electron dense accretion is a thin layer of cytoplasm which contains the nuclei. The outer surface of the resting sporangium is smooth. 相似文献
9.
Summary An ultrastructural study of the development of the resting sporangium ofSynchytrium endobioticum (Schilb.) Perc. infecting potato cells is presented. The resting sporangium is found to have a single large, centrally placed nucleus with a prominent nucleolus through its entirein situ development. The cytoplasmic organization of the resting sporangium is further characterized by numerous membrane-bound lipid bodies and osmiophilic bodies. The latter have a characteristic sieve-like appearance, probably because certain storage components have been extracted during preparation for electron microscopy. Because of the similar location and appearance of these osmiophilic bodies it is suggested that they are identical to what has earlier (based on light microscopy) been described as chromatin granules; and the ultrastructural studies presented here show that nucleolar discharge which was described from light microscopic observations as leading to chromatin granules in the cytoplasm, and finally forming the nuclei of the zoospores (bally 1912,curtis 1921,percival 1910) simply does not occur.The appearance of dense fibrillar-like structures on the sporangial surface at an early stage of resting sporangium development ultrastructurally distinguishes the resting sporangium from the zoosporangium. The development of the layered portion of the thick sporangial wall is shown to be due to the fusion of vacuoles containing pre-made wall fibrils with the cell membrane. It is suggested that the inner compact wall layer which is essentially substructureless is formed by the membrane itself.The characteristic wings of the matureS. endobioticum resting sporangium originate from the potato host cell wall. Remnants of host cell organelles in the outermost layer of the resting sporangium wall show that degradation of the host cell cytoplasm contributes to wall formation of the parasite. 相似文献
10.
Summary
Geosiphon pyriforme represents a photoautotrophic endosymbiosis of aGlomus-like fungus with the cyanobacteriumNostoc punctiforme. The fungus forms unicellular bladders of up to 2 mm in length and 0.5 mm in diameter growing on the soil surface and harboring the endosymbioticNostoc filaments. The cyanobacteria are located in a compartment (the symbiosome) bordered by a host membrane. The space between this symbiosome membrane (SM) and theNostoc cell wall is filled with an about 30–40 nm thick layer of amorphous material, which is present also in the regions of the symbiosome where noNostoc filaments are located. At these sites the amorphous material consists of a 20–30 nm thick layer separating the SM. The region between the SM and the cyanobacterium is defined as symbiosome space (SS). Fungal bladders, hyphae and free livingNostoc were analyzed by affinity techniques as well as the material occurring in the SS. FITC-coupled lectins with sugar specificity to -D-mannosyl/-D-glucosyl (Con A), N-acetyl--D-glucosamine oligomers (WGA), -L-fucosyl (UEA-I), -D-galactosyl (RCA-120), -D-galactosyl (BS-I-B4), N-acetyl--D-galactosamine (HPA), and sialic acid (EBL) residues were tested. WGA binding and calcofluor white staining demonstrated that the bladder wall as well as the SS contain fibrillar chitin. Of the other lectins only Con A clearly labeled the symbiosome. On the contrary, the lectin binding properties of the slime produced by free livingNostoc-colonies indicate the presence of mannose, fucose, GalNAc, sialic acid, and galactose, while chitin or GlucNAc-oligomers could not be detected. The symbiosome was also investigated electron microscopically. WGA-gold binding confirmed the presence of chitin, while a slight PATAg reaction indicated some polysaccharidic molecules within the SS. Our results show that the amorphous material within the SS contains molecules typical of the fungal cell wall and suggest that the SM is related to the fungal plasma membrane. The applied lectins all bind to the hyphal surface, indicating a high molecular complexity. Mannosyl, -galactosyl, and sialic acid residues are strongly exposed at the outer cell wall layer, whereas GlucNAc, GalNAc, and -galactosyl residues seem to be present in smaller amounts. The symbiotic interface established between the fungus andNostoc inGeosiphon shows many similarities to that occurring between fungi and root cells in arbuscular mycorrhizas.Abbreviations AM
arbuscular mycorrhiza
- BS-I-B4
Bandeiraea simplicifolia lectin I isolectin B4
- CLSM
confocal laser scanning microscopy
- Con A
Concanavalin A
- EBL
elderberry bark lectin I
- FITC
fluorescein isothiocyanate
- HPA
Helix pomatia agglutinin
- PATAg
periodic acid-thiocarbohydrazide-Ag proteinate
- SM
symbiosome membrane
- SS
symbiosome space
- RCA-120
Ricinus communis agglutinin 120
- UEA-I
Ulex europaeus agglutinin I
- WGA
wheat germ agglutinin
Dedicated to Professor Dr. Peter Sitte at the occasion of his 65th birthday 相似文献
11.
Fluorescent antibody staining indicated differences in surface antigenicity in Anabaena azollae cells fresh from the leaf cavities of the fern, Azolla caroliniana, and algae which were isolated and subcultured from this fern. Such results suggest that either changes in antigenicity occur in this phycobiont during culturing or that isolation selects for an antigenically different mutant strain capable of in vitro growth.Non-Standard Abbreviations FA
fluorescent antibody staining
- PBS
phosphate buffered saline
- W
microwatt
- Anti-F
antiserum prepared against fresh cells
- Anti-N
antiserum prepared against Newton's culture
- FTTC
fluorescein isothiocyanate
To whom offprint requests should be sent 相似文献
12.
Summary Undifferentiated ordinary epidermal cells (ECs) ofVigna sinensis leaves possess straight anticlinal walls and cortical microtubules (Mts) scattered along them. At an early stage of EC differentiation cortical Mts adjacent to the above walls form bundles normal to the leaf plane, loosely interconnected through the cortical cytoplasm of the internal periclinal wall. At the upper ends of the Mt bundles, Mts fan out towards the external periclinal wall and form radial arrays. Mt bundles and radial arrays exhibit strict alternate disposition between neighbouring ECs. An identical reticulum of cellulose microfibril (CM) bundles is deposited outside the Mt bundles. Local wall pads rise at the junctions of anticlinal walls with the external periclinal one, where the CM bundles terminate. They display radial CMs fanning towards the external periclinal wall. The CM bundles and radial CM systems prevent local cell bulging, but allow it in the intervening wall areas. In particular, the radial CM systems dictate the pattern of EC waviness by favouring local tangential expansion of external periclinal wall. As a result, ECs obtain an undulate appearance. Constrictions in one EC correspond with protrusions of adjacent ECs. ECs affected by colchicine entirely lose their Mts and do not develop wavy walls, an observation substantiating the role of cortical Mts in EC morphogenesis.Abbreviations CM
cellulose microfibril
- DTT
dithiothreitol
- EC
epidermal cell
- MSB
microtubule stabilizing buffer
- Mt
microtubule
- PBS
phosphate buffered saline
- PMSF
phenylmethylsulfonyl fluoride 相似文献
13.
Summary Differentiated mesophyll cells ofTriticum aestivum (cv. Star) exhibit a lobed outline resembling tube-shaped balloons with almost regularly spaced constrictions. It was shown that these constrictions are probably the result of hoops of wall reinforcements laid down during early stages of cell expansion. It appears that these hoops prevent expansion in the corresponding regions and thus give rise to the peculiar cell shape. The comparatively thin cell walls of the bulges are uniformly reinforced after the lobed shape is established.By using immunofluorescence techniques a change in the pattern of cortical microtubule arrangement was observed which corresponded to the pattern of cell wall deposition. Discrete bands of microtubules were found beneath the sites of hoop reinforcement. These bands disintegrated during late stages of cell expansion with microtubules fanning out into the almost empty regions of the bulges.Abbreviations DMSO
dimethyl sulfoxid
- EGTA
ethylene glycol bis-(-aminoethyl ether) N,N,N,N-tetraacetic acid
- FITC
fluorescein isothiocyanat
- MSB
microtubule stabilizing buffer
- PBS
phosphate buffered saline
- PIPES
1,4-piperazine diethanesulfonic acid
- PMSF
phenylmethyl sulfonylfluoride 相似文献
14.
We have investigated the in vitro reassembly of the salt soluble, hydroxyproline rich, glycoproteins from the cell wall of Chlamydomonas reinhardii, into structured cell wall fragments. We have devised an assay which has been used to follow the reassembly of the unfractionated and fractionated (2BI and 2BII) cell wall glycoproteins. Reassembly has a pH optimum of 5, a temperature optimum of 20°C, and the final size of the reassembled fragments appears to be promoted by the minor component 2BI. Periodate oxidation experiments show that sugar residues, in particular mannose, are important for accurate reassembly. Using electron microscopy, the structure of the reassembled products has been elucidated, as have intermediate stages in the reassembly process.Abbreviations TRIS
Tris(hydroxymethyl)-methylamine
- SDS
Sodium dodecyl sulphate
- PAGE
Polyacrylamide gel electrophoresis
This is the fifth paper in a series entitled Structure composition and morphogenesis of the cell wall of Chlamydomonas reinhardii. The last paper in this series was Catt et al. (1976) 相似文献
15.
Wolfram Braune 《Archives of microbiology》1980,126(3):257-261
Electronmicroscopical investigations of light activated akinetes in different phases before outgrowth of the germinating cell showed two alterations in the akinete envelope, obviously in connection with the germination process. After induction of germination the akinetes show formation of an expanding more or less electron dense layer between the outer cell wall layer (outer membrane, LIV) and the condensed part of the akinete coat (the transformed sheath of the vegetative cell). Between this new formed layer and the mentioned part of the akinete coat thick laminar layers are deposited which contain alternately electron dense and electron transparent strata. The expanding layer is assumed to be a mucous layer which acts as swelling body causing, after bursting of the layered shell, the expulsion of the germinating cell in the manner characteristic for Anabaena variabilis. 相似文献
16.
The (13)glucanase of Basidiomycete QM 806 was used to prepare Saccharomyces cerevisiae and Candida utilis protoplasts. Plasma membranes isolated from S. cerevisiae contained a small amount of mannose and traces of glucose and ribose. Randomly distributed -mannan was detected by scanning electron microscopy at the surface of prefixed protoplasts using colloidal gold labelled with Concanavalin A as a marker. C. utilis protoplasts were also marked with anti-mannan antibodies. Again the distribution of mannan was random. This experiment indicated also that plasma membrane mannan has the same immunochemical determinants as cell wall mannan. It is hypothesized that mannan is mainly located in the outer layer of plasma membranes. 相似文献
17.
Summary Monoclonal antibody PCBC3, raised against stylar extracts fromNicotians, alata flowers, was deduced from enzyme-linked immunosorbent assays and inhibition of immuno-gold labelling on tissue sections to bind specifically to carbohydrate epitopes on arabinogalactan proteins (AGPs) but not to other arabinose-containing cell wall polysaccharides. When pollen grains ofN. tabacum were hydrated in fixative, PCBC3 bound to vesicles in the vicinity of the endoplasmic reticulum but, when grains were hydrated for 20 min in culture medium before fixation, binding was restricted to the plasma membrane. The generative-cell plasma membrane was also labelled in grains ofLycopersicon peruvianum. In pollen tubes ofN. tabacum grown in liquid culture, the AGPs detected by PCBC3 were located in several regions, including the plasma membrane, tubular-vesicular structures (plasmalemmasomes) at and under the plasma membrane, and multilamellar bodies within vacuoles, features generally associated with endocytosis. Labelling was not evident in secretory vesicles or the plasma membrane at the pollen-tube tip. The AGPs detected with PCBC3 were also present in pollen-tube walls, near the interface between the inner, callosic layer and the outer, fibrillar, pectic layer. Pollen tubes ofN. tabacum grown in medium lacking added CuSO4 produce a wall with an abnormally thickened fibrillar layer, and this layer was uniformly labelled with PCBC3. The disposition of wall AGPs thus changes in pollen tubes of different morphologies.Abbreviations AGP
arabinogalactan protein
- -L-Araf
-L-arabinofuranose
- ELISA
enzyme-linked immunosorbent assay
- MAb
monoclonal antibody
- PBS
phosphate-buffered saline 相似文献
18.
Summary The cytoskeleton in epithelial cells ofSpongilla lacustris is constructed of microtubules radiating from the nuclear region and terminating at the cell periphery as well as microfilaments forming a cortical layer beneath the plasma membrane and distinct fibers in the cytoplasmic matrix. Single frame analysis and in vivo labeling of mitochondria with Rh 123, endosomes or lysosomes with TRITC-BSA, endoplasmic reticulum (ER) with DiOC6 (3) and dictyosomes with C6-NBD-ceramide points to the microtubular system as a candidate for controlled cytoplasmic organization and active transport of these cell organelles. In epithelial cells with an intact microtubular system, mitochondria and endosomes or lysosomes show a regular shuttle movement between the nucleus and the cell periphery with a velocity of 1.3–1.4 m/s; the ER forms a more or less dynamic two-dimensional network in the entire cytoplasmic matrix, and dictyosomes are arranged in a ring-like manner around the nucleus. In epithelial cells treated with colchicine or colcemid, mitochondria and endosomes or lysosomes gather in the perinuclear region and become immobile; the ER accumulates near the cell center, whereas most dictyosomes distribute randomly over the whole cytoplasm. Transformation of the microfilament system with cytochalasin D has no influence on cell organelle distribution and dynamics but impedes cell locomotion and cell surface activities.Abbreviations BSA
bovine serum albumin
- C6-NBD-ceramide
6-[(N-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]amino)caproyl]sphingosine
- DiOC6(3)
3,3-dihydroxyloxacarbocyanine jodide
- DMSO
dimethylsulfoxide
- EGTA
ethylenediaminetetraacetic acid
- GTX
glycerol-Triton-X-100
- PBS
phosphate buffered saline
- PEG
polyethylene glycol
- PIPES
1,4-piperazine-N,N-bis-(2-ethanesulfomc) acid
- Rh 123
rhodamine 123
- TRITC
tetramethylrhodamine isothiocyanate 相似文献
19.
Summary The localization of galactosyl residues and lectin binding sites in mucilage and cell walls of the colony forming green algaCosmocladium saxonicum (Desmidiaceae) has been studied using fluorescent probes. In mucilaginous filaments, which are secreted through pores of the cell wall, and in the primary cell wall galactosyl residues in -bound configuration are exposed, as indicated by indirect immunofluorescence using antiserum to monogalactosyl diglyceride residues. Concanavalin A receptors are present mainly at the surface of the secondary cell wall, whereasRicinus communis agglutinin, type I, receptors are predominantly associated with mucilaginous connecting strands, which join adjacent cells within a colony. NoUlex europaeus agglutinin receptors were found. Application of the fluorochrome calcofluor white ST resulted in labeling both, the primary and the secondary cell wall. The data, obtained with the fluorescent probes were compared with those obtained by thin layer chromatography of hydrolysed mucilage.This work includes parts of a doctoral thesis of B. S. carried out under the supervision of Prof. Dr. M.Mix. 相似文献
20.
D. J. W. Moriarty 《Archives of microbiology》1979,120(3):191-193
Muramic acid has been detected in Prochloron with the aid of two different techniques. It was assayed by cleaving D-lactate from muramic acid and then reducing NAD with D-lactate dehydrogenase and measuring the NADH with bacterial luciferase. Gas-liquid chromatography of trimethylsilyl derivatives of cell extracts confirmed that muramic acid was present in about the quantity given by the D-lactate assay. The amount of muramic acid present was 1.7±0.2 g/mg dry weight or 1.3fg/m2 of cell surface. This suggests that the thickness of the peptidoglycan layer in Prochloron is similar to that in blue-green algae.Abbreviations
D-LDH
d-lactate dehydrogenase
- MA
muramic acid
- TMS
trimethylsilyl
- TLE
thin layer electrophoresis
- GLC
gas-liquid chromatography 相似文献