scholarly journals Protein electroporation of Cre recombinase into cultured Arabidopsis cells with an intact cell wall

2019 ◽  
Author(s):  
Yoshio Kato ◽  
Yuichi Furuhata ◽  
Ayako Sakai ◽  
Yoshio Kato
Keyword(s):  
Cell Wall ◽  
Intact Cell ◽  
Cre Recombinase ◽  
Arabidopsis Cells

scholarly journals A method using electroporation for the protein delivery of Cre recombinase into cultured Arabidopsis cells with an intact cell wall

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yuichi Furuhata ◽  
Ayako Sakai ◽  
Tomi Murakami ◽  
Mone Morikawa ◽  
Chikashi Nakamura ◽  
...  
Keyword(s):  
Cell Wall ◽  
Protein Delivery ◽  
Intact Cell ◽  
Cre Recombinase ◽  
Arabidopsis Cells

scholarly journals Even Visually Intact Cell Walls in Waterlogged Archaeological Wood Are Chemically Deteriorated and Mechanically Fragile: A Case of a 170 Year-Old Shipwreck

Molecules ◽  
2020 ◽  
Vol 25 (5) ◽  
pp. 1113 ◽  
Author(s):  
Liuyang Han ◽  
Xingling Tian ◽  
Tobias Keplinger ◽  
Haibin Zhou ◽  
Ren Li ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Intact Cell ◽  
Nitrogen Adsorption ◽  
Wet Chemistry ◽  
Archaeological Wood ◽  
Waterlogged Archaeological Wood ◽  
X Ray Scattering ◽  
Crystallite Structure ◽  
Cell Wall Mechanics

Structural and chemical deterioration and its impact on cell wall mechanics were investigated for visually intact cell walls (VICWs) in waterlogged archaeological wood (WAW). Cell wall mechanical properties were examined by nanoindentation without prior embedding. WAW showed more than 25% decrease of both hardness and elastic modulus. Changes of cell wall composition, cellulose crystallite structure and porosity were investigated by ATR-FTIR imaging, Raman imaging, wet chemistry, 13C-solid state NMR, pyrolysis-GC/MS, wide angle X-ray scattering, and N2 nitrogen adsorption. VICWs in WAW possessed a cleavage of carboxyl in side chains of xylan, a serious loss of polysaccharides, and a partial breakage of β-O-4 interlinks in lignin. This was accompanied by a higher amount of mesopores in cell walls. Even VICWs in WAW were severely deteriorated at the nanoscale with impact on mechanics, which has strong implications for the conservation of archaeological shipwrecks.

scholarly journals Identification of Components of the Sigma B Regulon in Listeria monocytogenes That Contribute to Acid and Salt Tolerance

2008 ◽  
Vol 74 (22) ◽  
pp. 6848-6858 ◽  
Author(s):  
F. Abram ◽  
E. Starr ◽  
K. A. G. Karatzas ◽  
K. Matlawska-Wasowska ◽  
A. Boyd ◽  
...  
Keyword(s):  
Cell Wall ◽  
Listeria Monocytogenes ◽  
Stress Tolerance ◽  
Intact Cell ◽  
Microscopic Analysis ◽  
Phenotypic Characterization ◽  
Carbon Utilization ◽  
Two Dimensional Gel Electrophoresis ◽  
Sigma B ◽  
The Impact

ABSTRACT Sigma B (σB) is an alternative sigma factor that controls the transcriptional response to stress in Listeria monocytogenes and is also known to play a role in the virulence of this human pathogen. In the present study we investigated the impact of a sigB deletion on the proteome of L. monocytogenes grown in a chemically defined medium both in the presence and in the absence of osmotic stress (0.5 M NaCl). Two new phenotypes associated with the sigB deletion were identified using this medium. (i) Unexpectedly, the strain with the ΔsigB deletion was found to grow faster than the parent strain in the growth medium, but only when 0.5 M NaCl was present. This phenomenon was independent of the carbon source provided in the medium. (ii) The ΔsigB mutant was found to have unusual Gram staining properties compared to the parent, suggesting that σB contributes to the maintenance of an intact cell wall. A proteomic analysis was performed by two-dimensional gel electrophoresis, using cells growing in the exponential and stationary phases. Overall, 11 proteins were found to be differentially expressed in the wild type and the ΔsigB mutant; 10 of these proteins were expressed at lower levels in the mutant, and 1 was overexpressed in the mutant. All 11 proteins were identified by tandem mass spectrometry, and putative functions were assigned based on homology to proteins from other bacteria. Five proteins had putative functions related to carbon utilization (Lmo0539, Lmo0783, Lmo0913, Lmo1830, and Lmo2696), while three proteins were similar to proteins whose functions are unknown but that are known to be stress inducible (Lmo0796, Lmo2391, and Lmo2748). To gain further insight into the role of σB in L. monocytogenes, we deleted the genes encoding four of the proteins, lmo0796, lmo0913, lmo2391, and lmo2748. Phenotypic characterization of the mutants revealed that Lmo2748 plays a role in osmotolerance, while Lmo0796, Lmo0913, and Lmo2391 were all implicated in acid stress tolerance to various degrees. Invasion assays performed with Caco-2 cells indicated that none of the four genes was required for mammalian cell invasion. Microscopic analysis suggested that loss of Lmo2748 might contribute to the cell wall defect observed in the ΔsigB mutant. Overall, this study highlighted two new phenotypes associated with the loss of σB. It also demonstrated clear roles for σB in both osmotic and low-pH stress tolerance and identified specific components of the σB regulon that contribute to the responses observed.

Poly(β-hydroxybutyrate) extrusion from pleomorphic cells ofAzotobacter vinelandiiUWD

1995 ◽  
Vol 41 (13) ◽  
pp. 22-31 ◽  
Author(s):  
William J. Page ◽  
Luis D'elia ◽  
Richard Sherburne ◽  
Lori L. Graham
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Wall ◽  
Azotobacter Vinelandii ◽  
Intact Cell ◽  
Viable Cell ◽  
Transmission Electron ◽  
Polymer Expansion ◽  
Chemical Dehydration ◽  
Pleomorphic Cells

Azotobacter vinelandii UWD cells fill with up to 80% (per dry mass) poly(β-hydroxybutyrate) (PHB) after 24 h growth in medium containing sugars and fish peptone. However, peptones were not usually added to Azotobacter culture as they induced pleomorphism and compromised cell wall strength. This study examines the morphology of these PHB-producing pleomorphic cells in the transmission electron microscope. PHB-producing cells incubated for 18–24 h were most frequently 2–3 μm diameter spheres containing up to 20 PHB inclusions/cross section, or a calculated ≈ 100 inclusions/cell volume. These inclusions tended to be of small size (≈ 0.5 μm diameter) and became fewer and larger in older cells. The most striking feature of these pleomorphic cells was the apparent extrusion of polymer from the cells. It is unlikely that PHB extrusion is an active process from a viable cell as there was considerable cell wall damage at the point of polymer extrusion. The results suggest that the extrusion of PHB may be the result of polymer expansion, caused by the dehydration of the specimen for transmission electron microscopy, coupled with the inability of the pleomorphic cell wall to retain the expanding polymer. Thus, freeze-substituted sections of similar cells that were prepared without chemical dehydration did not extrude PHB. However, lysed cells prepared for transmission electron microscopy by chemical dehydration also did not extrude PHB, which suggests differences in the fluidity of the PHB in intact cell inclusions and lysed cell granules.Key words: poly(β-hydroxybutyrate), inclusions, polymer expansion, dehydration artifact.

A SEROLOGICAL STUDY OF CELL WALLS OF CERTAIN LACTIC ACID BACTERIA

1962 ◽  
Vol 8 (5) ◽  
pp. 629-637
Author(s):  
K. L. Chung ◽  
Roma Z. Hawirko
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Intact Cell ◽  
Synthetic Medium ◽  
Specific Cell ◽  
Intact Cells ◽  
Common Antigens ◽  
Species Specific ◽  
Relationship Of ◽  
The Relationship

From three species of Lactobacillus and three species of Streptococcus, cultured in a synthetic medium, cell walls were isolated following sonic disintegration and purified by washing. Sera against each species were prepared by injecting three rabbits with cell walls, and three with intact cells. Reciprocal agglutination tests were carried out with unabsorbed and absorbed antisera. More kinds of antibodies were detected with cell-wall antisera than with intact-cell antisera. Many species in the two genera shared common antigens. S. faecalis was the exception. Certain antigens believed to be complex haptens in nature reacted with heterologous antisera. Haemagglutination of tanned erythrocytes sensitized with a particulate cell-wall suspension showed fewer cross reactions than agglutination of intact-cell suspensions.The evidence presented shows the possibility of using antisera against species-specific cell-wall antigens for the identification of these species. The relationship of these species is discussed.

scholarly journals The Chlamydomonas cell wall and its constituent glycoproteins analyzed by the quick-freeze, deep-etch technique.

1985 ◽  
Vol 101 (4) ◽  
pp. 1550-1568 ◽  
Author(s):  
U W Goodenough ◽  
J E Heuser
Keyword(s):  
Cell Wall ◽  
Intact Cell ◽  
Experimental Conditions ◽  
Fibrous Matrix ◽  
Additional Information ◽  
Fibrous Network ◽  
Innermost Layer ◽  
Flagellar Membrane ◽  
Sexual Agglutinins ◽  
Mechanical Shearing

Using the quick-freeze, deep-etch technique, we have analyzed the structure of the intact cell wall of Chlamydomonas reinhardi, and have visualized its component glycoproteins after mechanical shearing and after depolymerization induced by perchlorate or by the wall-disrupting agent, autolysin. The intact wall has previously been shown in a thin-section study (Roberts, K., M. Gurney-Smith, and G. J. Hills, 1972, J. Ultrastruct. Res. 40:599-613) to consist of a discrete central triplet bisecting a meshwork of fibrils. The deep-etch technique provides additional information about the architecture of each of these layers under several different experimental conditions, and demonstrates that each layer is constructed from a distinct set of components. The innermost layer of the central triplet proves to be a fibrous network which is stable to perchlorate but destabilized by autolysin, disassembling into fibrillar units we designate as "fishbones." The medial layer of the triplet is a loose assemblage of large granules. The outer layer is a thin, crystalline assembly that is relatively unaffected by autolysin. It depolymerizes into two glycoprotein species, one fibrous and one globular. The wall glycoproteins prove to be structurally similar to two fibrous proteins that associate with the flagellar membrane, namely, the sexual agglutinins and the protomers of a structure we designate a "hammock." They are also homologous to some of the fibrous components found in the extracellular matrices of multicellular plants and animals. The quick-freeze, deep-etch technique is demonstrated to be a highly informative way to dissect the structure of a fibrous matrix and visualize its component macromolecules.

scholarly journals The role of plant cell wall encapsulation and porosity in regulating lipolysis during the digestion of almond seeds

2016 ◽  
Vol 7 (1) ◽  
pp. 69-78 ◽  
Author(s):  
Myriam M. L. Grundy ◽  
Frédéric Carrière ◽  
Alan R. Mackie ◽  
David A. Gray ◽  
Peter J. Butterworth ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Intact Cell ◽  
Lipid Digestion

Intact cell walls of almond prevent lipase penetration thus hindering lipid digestion.

Studies on the cell wall of Spirillum serpens. 1. Isolation and partial purification of the outermost cell wall layer

1970 ◽  
Vol 16 (10) ◽  
pp. 1011-1022 ◽  
Author(s):  
Francis L. A. Buckmire ◽  
Robert G. E. Murray
Keyword(s):  
Cell Wall ◽  
Intact Cell ◽  
Guanidine Hydrochloride ◽  
Casein Hydrolysate ◽  
Wall Layer ◽  
Partial Purification ◽  
Repeated Heating ◽  
Physical Study ◽  
Backing Layer ◽  
Wall Components

The presence of the hexagonal array of macromolecules on the outer surface of the cell wall of Spirillum serpens VHA required the addition of calcium to an otherwise effective growth medium (vitamin-free casein hydrolysate); slightly improved growth resulted from addition of a complex salts mixture. A means of isolating this layer for chemical and physical study was sought and controlled by electron microscopy of freeze-etched, negatively stained, and sectioned preparations. The structure was destroyed, extracted, or removed by extremes of pH (<4.5, >9), 1 M guanidine hydrochloride (pH 7), 2 M urea, and dimethyl sulfoxide, with varying damage to the cell. Heat (60° for 1 h) removed much of the outer layer from the intact cell as an array of units disposed on a delicate backing layer, leaving the basic Gram-negative triplet wall components. These fragments remained stable through washing and repeated heating in the presence of 0.001 M calcium chloride. Guanidine hydrochloride (1.5 M) dissolved the units from the tubes and vesicles formed by the backing layer. Dialysis against water removed salts and guanidine, caused the precipitation of residual contaminants, and provided a supernatant which, when lyophilized, provided a product containing 98% protein.

Topographic Features of Neisseria Catarrhalis Strains

1973 ◽  
Vol 31 ◽  
pp. 572-573
Author(s):  
G. A. Wistreich ◽  
R. F. Baker ◽  
W. B. Hammond ◽  
D. Ivler
Keyword(s):  
Biological Sciences ◽  
Cell Wall ◽  
Los Angeles ◽  
Intact Cell ◽  
Bacterial Species ◽  
Culture Collection ◽  
Surface Pattern ◽  
University Of Southern California ◽  
Topographic Features ◽  
Wall Structures

The surface features of several bacterial species belonging to the Neisseriaceae including Neisseria gonorrhoeae, N. meningitidis and Veillonella parvula have been reported. During fimbriation studies with N. catarrhalis negative staining revealed a surface pattern differing from those found with other species of this genus. This report presents a comparative study with three N. catarrhalis strains. Consideration is given only to the physical aspects of intact cell wall structures.Of the three Neisseria catarrhalis strains used, one came from the stock culture collection of the Bacteriology Section, Division of Biological Sciences, University of Southern California, Los Angeles, while the remaining strains (ATCC 8176, ATCC 8193), were from the American Type Culture Collection, Rockville, Maryland.

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