Use of a DNA probe for mapping by electron microscopy the ribosomal sequences in ribosomal RNA precursors from duck cells

1974 ◽  
Vol 90 (1) ◽  
pp. 65-76 ◽  
Author(s):  
William I. Murphy ◽  
Giuseppe Attardi
Keyword(s):  
Electron Microscopy ◽  
Ribosomal Rna ◽  
Dna Probe

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

scholarly journals Synthesis of a biotinylated DNA probe to detect ribosomal RNA cistrons inProvidencia stuartii

1987 ◽  
Vol 48 (1-2) ◽  
pp. 283-287 ◽  
Author(s):  
D.G. Pitcher ◽  
R.J. Owen ◽  
P. Dyal ◽  
A. Beck
Keyword(s):  
Ribosomal Rna ◽  
Dna Probe

Identification and Characterization of Salmonella Isolates by Automated Ribotyping†

1998 ◽  
Vol 61 (5) ◽  
pp. 519-524 ◽  
Author(s):  
THOMAS P. OSCAR
Keyword(s):  
Ribosomal Rna ◽  
Dna Probe ◽  
Strain Level ◽  
Reference Laboratory ◽  
E Coli ◽  
Genus Level ◽  
Typing Methods ◽  
Automated Ribotyping ◽  
Identification And Characterization

A study was conducted with the RiboPrinter, an automated ribotyping system, to evaluate its ability to identify and characterize isolates of Salmonella from broiler operations. Isolates of Salmonella obtained from a local broiler company were serotyped by a reference laboratory and ribotyped using the RiboPrinter. The RiboPrinter generated ribotype patterns by probing EcoRI digests of Salmonella DNA with an E. coli DNA probe to the ribosomal RNA operon. The RiboPrinter identified isolates by band matching of their ribotype patterns to ribotype patterns in its database. In addition, the RiboPrinter characterized isolates by sorting them into ribotypes on the basis of the similarity of their ribotype patterns. Of 117 isolates, the RiboPrinter identified 34 (29%) at the serotype level, 11 (9%) at the strain level, 46 (39%) at the genus level, and 26 (22%) were not identified. Thus, only 38% of the isolates were identified at or below the serotype level, indicating that the RiboPrinter was limited in its ability to identify Salmonella isolates by band matching. In contrast, the RiboPrinter was very effective at characterizing Salmonella isolates. Out of 108 isolates, the RiboPrinter detected 31 ribotypes, compared to serotyping which only detected 22 types of Salmonella. Thus, automated ribotyping was more discriminatory than serotyping. However, when results of both typing methods were combined, 40 types of Salmonella were detected, indicating that the best discrimination was obtained when automated ribotyping and serotyping were used together.

Electron Microscopy Analysis of the Nucleolus ofTrypanosoma cruzi

2005 ◽  
Vol 11 (4) ◽  
pp. 293-299 ◽  
Author(s):  
Gabriel López-Velázquez ◽  
Roberto Hernández ◽  
Imelda López-Villaseñor ◽  
Horacio Reyes-Vivas ◽  
María de L. Segura-Valdez ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Ribosomal Rna ◽  
Three Dimensional ◽  
Nuclear Bodies ◽  
Morphological Data ◽  
Central Position ◽  
Main Site ◽  
Dimensional Reconstruction ◽  
Electron Microscopy Analysis ◽  
Synthesis And Processing

The nucleolus is the main site for synthesis and processing of ribosomal RNA in eukaryotes. In mammals, plants, and yeast the nucleolus has been extensively characterized by electron microscopy, but in the majority of the unicellular eukaryotes no such studies have been performed. Here we used ultrastructural cytochemical and immunocytochemical techniques as well as three-dimensional reconstruction to analyze the nucleolus ofTrypanosoma cruzi, which is an early divergent eukaryote of medical importance. InT. cruziepimastigotes the nucleolus is a spherical intranuclear ribonucleoprotein organelle localized in a relatively central position within the nucleus. Dense fibrillar and granular components but not fibrillar centers were observed. In addition, nuclear bodies resembling Cajal bodies were observed associated to the nucleolus in the surrounding nucleoplasm. Our results provide additional morphological data to better understand the synthesis and processing of the ribosomal RNA in kinetoplastids.

Structure of partially denatured Escherichia coli 23 S ribosomal RNA determined by electron microscopy

1983 ◽  
Vol 168 (4) ◽  
pp. 809-830 ◽  
Author(s):  
Barbara Kure Klein ◽  
Thomas C. King ◽  
David Schlessinger
Keyword(s):  
Electron Microscopy ◽  
Escherichia Coli ◽  
Ribosomal Rna

scholarly journals Structure of the eukaryotic cytoplasmic pre-40S ribosomal subunit

2017 ◽  
Author(s):  
Alain Scaiola ◽  
Cohue Peña ◽  
Melanie Weisser ◽  
Daniel Böhringer ◽  
Marc Leibundgut ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Ribosomal Rna ◽  
Ribosomal Subunit ◽  
Mature Form ◽  
Structural Framework ◽  
Cryo Electron Microscopy ◽  
40S Ribosomal Subunit ◽  
Final Maturation ◽  
Assembly Factors ◽  
Conformational Rearrangements

AbstractFinal maturation of eukaryotic ribosomes occurs in the cytoplasm and requires the sequential removal of associated assembly factors and processing of the immature 20S pre-RNA. Using cryo-electron microscopy (cryo-EM), we have determined the structure of a cytoplasmic pre-40S particle poised to initiate final maturation at a resolution of 3.4 Å. The structure reveals the extent of conformational rearrangements of the 3’ major and 3’ minor domains of the ribosomal RNA that take place during maturation, as well as the roles of the assembly factors Enp1, Ltv1, Rio2, Tsr1, and Pno1 in the process. Altogether, we provide a structural framework for the coordination of the final maturation events that drive a pre-40S particle towards the mature form capable of engaging in translation.

scholarly journals A study of the effects of reaction with formaldehyde on some optical and physical properties of reticulocyte ribosomes

1969 ◽  
Vol 114 (4) ◽  
pp. 743-751 ◽  
Author(s):  
R A Cox
Keyword(s):  
Electron Microscopy ◽  
Absorption Spectrum ◽  
Ribosomal Rna ◽  
Sodium Dodecyl Sulphate ◽  
Sodium Dodecyl ◽  
Sedimentation Coefficient ◽  
Optical Rotatory Dispersion ◽  
Ultraviolet Absorption Spectrum ◽  
Optical Rotatory

1. The optical rotatory dispersion and ultraviolet-absorption spectrum of ribosomal RNA in situ appear to be unchanged when the ribosome is dissociated into its RNA and protein moieties. 2. Reaction with 0·05% formaldehyde at 20° for 2hr. ‘fixes’ ribosomes so that they remain intact in 1% sodium dodecyl sulphate. 3. The RNA moiety of the ribosome undergoes a conformational change when ribosomes in 8% formaldehyde are heated at 70° for 10min. and cooled to 20°. After this treatment no double-helical character can be detected, but neither the sedimentation coefficient nor the morphology of the ribosome determined by electron microscopy is altered. 4. It is concluded that the RNA moiety of reticulocyte ribosomes is freely accessible to formaldehyde.

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