蓖麻毒素、志贺毒素和相关核糖体失活蛋白的功能性定量聚合酶链反应检测法。

A functional quantitative polymerase chain reaction assay for ricin, Shiga toxin, and related ribosome-inactivating proteins.

机构信息

Division of Biochemical Toxicology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA.

出版信息

Anal Biochem. 2010 Jan 15;396(2):204-11. doi: 10.1016/j.ab.2009.09.024. Epub 2009 Sep 17.

DOI:10.1016/j.ab.2009.09.024

PMID:19766090

Abstract

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).

摘要

蓖麻毒素、相思豆毒素和其他核糖体失活蛋白等强效毒素使 28S 核糖体 RNA 中的一个特定碱基失活，从而破坏核糖体并导致细胞死亡。我们利用逆转录酶优先在 RNA 中的无碱基位点对面插入腺嘌呤这一事实，创建了一种定量聚合酶链反应（PCR）检测法来检测损伤。该检测法可检测到低至 30pg 的蓖麻毒素。我们使用该检测法研究了蓖麻毒素的酶促特性，如最适 pH 值和温度（pH4.5-5.0 和 60°C）。