scholarly journals Heterologous Expression and High Degree Purification of the Restriction Endonuclease SauUSI

BIO-PROTOCOL ◽  
2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Vinayak Tumuluri ◽  
Kayarat Saikrishnan
Keyword(s):  
Heterologous Expression ◽  
Restriction Endonuclease ◽  
High Degree

Molecular analyses of conjugative, gentamicin-resistance plasmids from staphylococcal clinical isolates

1988 ◽  
Vol 34 (9) ◽  
pp. 1050-1057 ◽  
Author(s):  
Ioannis Zorbas ◽  
Robert T. Hall ◽  
Sue L. Hall ◽  
William G. Barnes ◽  
Marvin Rogolsky
Keyword(s):  
Restriction Endonuclease ◽  
Clinical Isolates ◽  
Genetic Determinants ◽  
Conjugative Plasmids ◽  
Molecular Weights ◽  
Geographical Regions ◽  
Gentamicin Resistance ◽  
Ecori Restriction ◽  
High Degree ◽  
Endonuclease Digestion

Gentamicin-resistant Staphylococcus aureus and Staphylococcus epidermidis strains which were isolated from infants with staphylococcal bacteremia were analyzed for the presence of self-transmissible gentamicin-resistance (Gmr) plasmids. Conjugative GMr plasmids of approximately 43.8–63 kilobases (kb) were found in all S. aureus strains. Inter- and intra-species transfer of Gmr plasmids by conjugation was observed from S. aureus to S. aureus and to S. epidermidis recipient strains. However, neither inter- nor intra-species transfer of gentamicin resistance by conjugation was observed with nine out of nine S. epidermidis donor strains which were mated with either S. epidermidis or S. aureus recipient strains. These conjugative Gmr plasmids were unable to comobilize a smaller (15-kb) plasmid present in all but two S. aureus clinical isolates. Many of the conjugative Gmr plasmids also carried genetic determinants for kanamycin, tobramycin, neomycin, and ethidium bromide resistance, and for β-lactamase synthesis. EcoRI restriction endonuclease digests of the S. aureus Gmr conjugative plasmids revealed three different digestion patterns. Four EcoRI restriction endonuclease digestion fragments of 15, 11.4, 6.3, and 4.6 kb in size were common to all plasmids. These plasmids and conjugative Gmr staphylococcal plasmids from other geographical regions shared restriction digestion fragments of similar molecular weights. DNA hybridization with biotinylated S. aureus plasmid pIZ7814 DNA revealed a high degree of homology among these plasmids. A 50.9-kb plasmid from one of the nonconjugative S. epidermidis clinical isolates showed homology with the probe DNA but lacked a portion of a 6.3-kb fragment which was present in all conjugative plasmids and believed to carry much genetic information for conjugation.

Predominance of a Single Restriction Endonuclease Analysis Group with Intrahospital Subgroup Diversity AmongClostridium difficileIsolates at Two Chicago Hospitals

2002 ◽  
Vol 23 (11) ◽  
pp. 648-652 ◽  
Author(s):  
Endale T. Mekonen ◽  
Dale N. Gerding ◽  
Susan P. Sambol ◽  
Jean M. Pottinger ◽  
Joseph J. Pulvirenti ◽  
...  
Keyword(s):  
Restriction Endonuclease ◽  
Clinical Laboratory ◽  
Tertiary Care ◽  
Restriction Endonuclease Analysis ◽  
Veterans Affairs ◽  
County Hospital ◽  
Analysis Group ◽  
High Degree ◽  
Tertiary Care Hospitals ◽  
Endonuclease Analysis

Objective:To determine the epidemiology and relatedness ofClostridium difficileisolates in two geographically separated hospitals in a large metropolitan area, each with unique patients and personnel.Design:Observational descriptive molecular epidemiology of clinicalC. difficileisolates.Setting:Two tertiary-care hospitals in Chicago.Methods:ConsecutiveC. difficileisolates from the clinical laboratory of a Veterans Affairs hospital during a 13-month period were typed by restriction endonuclease analysis (REA). During an overlapping 3-month period, stool specimens that tested positive forC. difficiletoxin from patients at a nearby county hospital were cultured and the recovered isolates typed by the same method.Results:Nineteen (68%) of 28 nosocomial isolates at the smaller, Veterans Affairs hospital belonged to REA group K. Within this group of closely related strains, 9 distinct REA types were recognized. Twenty-one (72%) of 29 nosocomial isolates at the larger, county hospital also belonged to group K. However, the predominant REA types within group K differed markedly at each institution.Conclusions:These findings demonstrate a high degree of similarity among nosocomialC. difficilestrains from different hospitals in the same city and suggest the possibility of an extended outbreak of a prototype group K strain with subsequent genetic drift at the two different institutions.

scholarly journals Studies on the heterologous expression of BstVI restriction endonuclease in Escherichia coli

IUBMB Life ◽  
1998 ◽  
Vol 44 (2) ◽  
pp. 391-397
Author(s):  
Claudia Saavedra ◽  
Enrique González ◽  
Claudio Vásquez
Keyword(s):  
Escherichia Coli ◽  
Heterologous Expression ◽  
Restriction Endonuclease

Tissue section lifting for electron microscopy

1978 ◽  
Vol 36 (2) ◽  
pp. 86-87
Author(s):  
Adrian F. van Dellen
Keyword(s):  
Infectious Disease ◽  
Electron Microscopy ◽  
Tissue Culture ◽  
Organ System ◽  
Surrounding Tissue ◽  
Paraffin Sections ◽  
Surface Areas ◽  
Specific Determination ◽  
High Degree ◽  
Accuracy And Repeatability

The morphologic pathologist may require information on the ultrastructure of a non-specific lesion seen under the light microscope before he can make a specific determination. Such lesions, when caused by infectious disease agents, may be sparsely distributed in any organ system. Tissue culture systems, too, may only have widely dispersed foci suitable for ultrastructural study. In these situations, when only a few, small foci in large tissue areas are useful for electron microscopy, it is advantageous to employ a methodology which rapidly selects a single tissue focus that is expected to yield beneficial ultrastructural data from amongst the surrounding tissue. This is in essence what "LIFTING" accomplishes. We have developed LIFTING to a high degree of accuracy and repeatability utilizing the Microlift (Fig 1), and have successfully applied it to tissue culture monolayers, histologic paraffin sections, and tissue blocks with large surface areas that had been initially fixed for either light or electron microscopy.

Electron Microscopy in Molecular Biology

1967 ◽  
Vol 25 ◽  
pp. 2-3
Author(s):  
Cecil E. Hall
Keyword(s):  
Electron Microscopy ◽  
Molecular Biology ◽  
Noise Level ◽  
Molecular Structures ◽  
Material Structure ◽  
The Arts ◽  
Shadow Casting ◽  
Electron Image ◽  
High Degree ◽  
Very High

The visualization of organic macromolecules such as proteins, nucleic acids, viruses and virus components has reached its high degree of effectiveness owing to refinements and reliability of instruments and to the invention of methods for enhancing the structure of these materials within the electron image. The latter techniques have been most important because what can be seen depends upon the molecular and atomic character of the object as modified which is rarely evident in the pristine material. Structure may thus be displayed by the arts of positive and negative staining, shadow casting, replication and other techniques. Enhancement of contrast, which delineates bounds of isolated macromolecules has been effected progressively over the years as illustrated in Figs. 1, 2, 3 and 4 by these methods. We now look to the future wondering what other visions are waiting to be seen. The instrument designers will need to exact from the arts of fabrication the performance that theory has prescribed as well as methods for phase and interference contrast with explorations of the potentialities of very high and very low voltages. Chemistry must play an increasingly important part in future progress by providing specific stain molecules of high visibility, substrates of vanishing “noise” level and means for preservation of molecular structures that usually exist in a solvated condition.

A High Angle, Double Tilting Cold Stage for the AEI EM7

1974 ◽  
Vol 32 ◽  
pp. 450-451
Author(s):  
P.R. Swann ◽  
A.E. Lloyd
Keyword(s):  
Habit Plane ◽  
Temperature Control ◽  
Tilt Angle ◽  
Cooling System ◽  
Thermal Drift ◽  
High Angle ◽  
Cold Stage ◽  
High Degree ◽  
Better Than

Figure 1 shows the design of a specimen stage used for the in situ observation of phase transformations in the temperature range between ambient and −160°C. The design has the following features a high degree of specimen stability during tilting linear tilt actuation about two orthogonal axes for accurate control of tilt angle read-out high angle tilt range for stereo work and habit plane determination simple, robust construction temperature control of better than ±0.5°C minimum thermal drift and transmission of vibration from the cooling system.

New Aspects in the Design of Electron Optical Magnification Systems

1972 ◽  
Vol 30 ◽  
pp. 606-607
Author(s):  
Willem H.J. Andersen
Keyword(s):  
Electron Microscope ◽  
Imaging System ◽  
Chromatic Aberration ◽  
Research Tool ◽  
Energy Losses ◽  
Objective Lens ◽  
Theoretical Limit ◽  
Optical Magnification ◽  
Chromatic Aberrations ◽  
High Degree

Electron microscope design, and particularly the design of the imaging system, has reached a high degree of perfection. Present objective lenses perform up to their theoretical limit, while the whole imaging system, consisting of three or four lenses, provides very wide ranges of magnification and diffraction camera length with virtually no distortion of the image. Evolution of the electron microscope in to a routine research tool in which objects of steadily increasing thickness are investigated, has made it necessary for the designer to pay special attention to the chromatic aberrations of the magnification system (as distinct from the chromatic aberration of the objective lens). These chromatic aberrations cause edge un-sharpness of the image due to electrons which have suffered energy losses in the object.There exist two kinds of chromatic aberration of the magnification system; the chromatic change of magnification, characterized by the coefficient Cm, and the chromatic change of rotation given by Cp.

Cytoplasmic Organization in the Receptor Cells of the Crista Ampullaris

1972 ◽  
Vol 30 ◽  
pp. 60-61 ◽  
Author(s):  
Robert F. Dunn
Keyword(s):  
Fine Particles ◽  
Apical Cell ◽  
Apical Region ◽  
Receptor Cells ◽  
Cuticular Plate ◽  
Morphological Organization ◽  
Crista Ampullaris ◽  
Cytoplasmic Organization ◽  
High Degree ◽  
General Appearance

Receptor cells of the cristae in the vestibular labyrinth of the bullfrog, Rana catesbiana, show a high degree of morphological organization. Four specialized regions may be distinguished: the apical region, the supranuclear region, the paranuclear region, and the basilar region.The apical region includes a single kinocilium, approximately 40 stereocilia, and many small microvilli all projecting from the apical cell surface into the lumen of the ampulla. A cuticular plate, located at the base of the stereocilia, contains filamentous attachments of the stereocilia, and has the general appearance of a homogeneous aggregation of fine particles (Fig. 1). An accumulation of mitochondria is located within the cytoplasm basal to the cuticular plate.

Computer Reconstruction of the Cellular Architecture of the Daphnia Magna Optic Ganglion

1975 ◽  
Vol 33 ◽  
pp. 284-285
Author(s):  
E. R. Macagno ◽  
C. Levinthal
Keyword(s):  
Daphnia Magna ◽  
Genetic Background ◽  
Compound Eye ◽  
Synaptic Connectivity ◽  
Serial Sections ◽  
Cross Sectional ◽  
Small Crustacean ◽  
Optic Ganglion ◽  
Structural Invariance ◽  
High Degree

The optic ganglion of Daphnia Magna, a small crustacean that reproduces parthenogenetically contains about three hundred neurons: 110 neurons in the Lamina or anterior region and about 190 neurons in the Medulla or posterior region. The ganglion lies in the midplane of the organism and shows a high degree of left-right symmetry in its structures. The Lamina neurons form the first projection of the visual output from 176 retinula cells in the compound eye. In order to answer questions about structural invariance under constant genetic background, we have begun to reconstruct in detail the morphology and synaptic connectivity of various neurons in this ganglion from electron micrographs of serial sections (1). The ganglion is sectioned in a dorso-ventra1 direction so as to minimize the cross-sectional area photographed in each section. This area is about 60 μm x 120 μm, and hence most of the ganglion fit in a single 70 mm micrograph at the lowest magnification (685x) available on our Zeiss EM9-S.

Feasibility of Molecular Structure Determination With a High Resolution Electron Microscope

1968 ◽  
Vol 26 ◽  
pp. 338-339
Author(s):  
T. A. Welton
Keyword(s):  
Electron Microscope ◽  
Substrate Surface ◽  
Helical Axis ◽  
Base Plane ◽  
Resolution Electron ◽  
High Resolution Electron Microscope ◽  
High Degree ◽  
Degree Of Order ◽  
Purine Pyrimidine

An ultimate design goal for an improved electron microscope, aimed at biological applications, is the determination of the structure of complex bio-molecules. As a prototype of this class of problems, we propose to examine the possibility of reading DNA sequence by an imaginable instrument design. This problem ideally combines absolute importance and relative simplicity, in as much as the problem of enzyme structure seems to be a much more difficult one.The proposed technique involves the deposition on a thin graphite lamina of intact double helical DNA rods. If the structure can be maintained under vacuum conditions, we can then make use of the high degree of order to greatly reduce the work involved in discriminating between the four possible purine-pyrimidine arrangements in each base plane. The phosphorus atoms of the back bone form in projection (the helical axis being necessarily parallel to the substrate surface) two intertwined sinusoids. If these phosphorus atoms have been located up to a certain point on the molecule, we have available excellent information on the orientation of the base plane at that point, and can then locate in projection the key atoms for discrimination of the four alternatives.

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