A functional quantitative polymerase chain reaction assay for ricin, Shiga toxin, and related ribosome-inactivating proteins
Autor: | William B. Melchior, William H. Tolleson |
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Rok vydání: | 2010 |
Předmět: |
Ribosome Inactivating Proteins
Biophysics Apoptosis Ricin Biology Polymerase Chain Reaction Biochemistry Ribosome Cell Line Shiga Toxin chemistry.chemical_compound Animals Molecular Biology Ribosome-inactivating protein Temperature RNA RNA-Directed DNA Polymerase Shiga toxin Cell Biology Hydrogen-Ion Concentration Molecular biology Reverse transcriptase Rats carbohydrates (lipids) enzymes and coenzymes (carbohydrates) Real-time polymerase chain reaction chemistry biology.protein Abrin |
Zdroj: | Analytical Biochemistry. 396:204-211 |
ISSN: | 0003-2697 |
DOI: | 10.1016/j.ab.2009.09.024 |
Popis: | The potent toxins ricin, abrin, and other ribosome-inactivating proteins deadenylate a specific base in 28S ribosomal RNA that destroys ribosomes and leads to cell death. We have taken advantage of the fact that reverse transcriptase preferentially inserts an adenine opposite to an abasic site in RNA to create a quantitative polymerase chain reaction (PCR) assay to detect the damage. This assay detects as little as 30pg of ricin. We used the assay to study enzymatic properties of ricin such as pH and temperature optima (pH 4.5-5.0 and 60 degrees C). |
Databáze: | OpenAIRE |
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