University of California

Bruce E Lehnert

Paul J Jackson

John P Nolan

Rhiannon L Nolan

Nick M Cirino

Teresa Ruscetti

The protective antigen (PA) of,is integral to the mechanism of anthrax poisoning. The cloning, expression and purification of a 32 kDa,PA fragment (PA32) is described. This fragment has also been expressed as a fusion construct to stabilized green fluorescent protein (EGFP-PA32). Both proteins were capable of binding to specific cell surface receptors as determined by fluorescent microscopy and a flow cytometric assay. To confirm binding specificity in the flow cytometric assay, non-fluorescent PA83 or PA32 was used to competitively inhibit fluorescent EGFP-PA32 binding to cell receptors. This assay can be employed as a rapid screen for compounds which disrupts binding of PA to cells. Additionally, the high intracellular expression levels and ease of purification make this recombinant protein an attractive vaccine candidate or therapeutic treatment for anthrax poisoning.

Method for screening inhibitors of the toxicity of Bacillus anthracis

A method for making recombinant fluorescent protein standard particles for calibration of fluorescence instruments.

Growing community of inventors

Los Alamos, NM, United States of America

Bruce E Lehnert