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用于实时核酸酶动力学的定量微孔板检测法

Quantitative Microplate Assay for Real-Time Nuclease Kinetics.

作者信息

Eriksson Jonas, Langel Ülo

机构信息

Department of Neurochemistry, Stockholm University, Stockholm, Sweden.

出版信息

PLoS One. 2016 Apr 21;11(4):e0154099. doi: 10.1371/journal.pone.0154099. eCollection 2016.

DOI:10.1371/journal.pone.0154099
PMID:27101307
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4839650/
Abstract

Utilizing the phenomenon of nucleases exposing oligonucleotide phosphate backbones to phosphatases we present a novel quantitative method for kinetics of nuclease catalysis. Inorganic phosphate released from nuclease products by phosphatases could be quantified in real-time by a fluorescent sensor of inorganic phosphate. Two different nucleases were employed, showing the versatility of this assay for multiple turnover label-free nuclease studies.

摘要

利用核酸酶将寡核苷酸磷酸骨架暴露给磷酸酶的现象，我们提出了一种用于核酸酶催化动力学的新型定量方法。磷酸酶从核酸酶产物中释放的无机磷酸盐可以通过无机磷酸盐荧光传感器进行实时定量。我们使用了两种不同的核酸酶，这表明该检测方法在多种周转无标记核酸酶研究中的通用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/4f9e3813c2c1/pone.0154099.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/229074cd7146/pone.0154099.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/b23bee376197/pone.0154099.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/3f608d3087ef/pone.0154099.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/4f9e3813c2c1/pone.0154099.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/229074cd7146/pone.0154099.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/b23bee376197/pone.0154099.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/3f608d3087ef/pone.0154099.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92d1/4839650/4f9e3813c2c1/pone.0154099.g004.jpg

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