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University of Virginia Health System

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Development of a Negative Selectable Marker for Entamoeba histolytica

Mayuresh M Abhyankar¹, Sarah M Haviland¹, Carol A Gilchrist¹, William A Petri, Jr.¹

¹Division of Infectious Disease and International Health, University of Virginia Health System

We report development of a negative selection system in E. histolytica based upon transgenic expression of a chimeric protein (FCU1) and selection with the prodrug 5-fluorocytosine. The FCU1 protein is a fusion of yeast cytosine deaminase and uracil phosphoribosyltransferase. Expression of FCU1 resulted in increased E. histolytica sensitivity towards 5-fluorocytosine.

Immunology and Infection

Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction

Michael R. Davis, Jr.¹, Joanna B. Goldberg¹

¹Department of Microbiology, Immunology, & Cancer Biology, University of Virginia Health System

We describe a modified hot aqueous-phenol extraction method for purifying lipopolysaccharide (LPS) from Gram-negative bacteria. Once extracted, the LPS can be subsequently analyzed by SDS-PAGE and visualized by direct staining or Western immunoblot.

Neuroscience

Preparation of Newborn Rat Brain Tissue for Ultrastructural Morphometric Analysis of Synaptic Vesicle Distribution at Nerve Terminals

Bianca Ferrarese^1,2, Nadia Lunardi¹

¹Department of Anesthesiology, University of Virginia Health System, ²Department of Anesthesiology, Universita' degli Studi di Padova

We describe procedures for processing newborn rat brain tissue to obtain high-resolution electron micrographs for morphometric analysis of synaptic vesicle distribution at nerve terminals. The micrographs obtained with these methods can also be used to study the morphology of a number of other cellular components and their dimensional structural relationships.