通过荧光相关光谱法研究病毒DNA包装
Viral DNA packaging studied by fluorescence correlation spectroscopy.
作者信息
Sabanayagam Chandran R, Oram Mark, Lakowicz Joseph R, Black Lindsay W
出版信息
Biophys J. 2007 Aug 15;93(4):L17-9. doi: 10.1529/biophysj.107.111526. Epub 2007 Jun 8.
Abstract
The DNA packaging machinery of bacteriophage T4 was studied in vitro using fluorescence correlation spectroscopy. The ATP-dependent translocation kinetics of labeled DNA from the bulk solution, to the phage interior, was measured by monitoring the accompanied decrease in DNA diffusibility. It was found that multiple short DNA fragments (100 basepairs) can be sequentially packaged by an individual phage prohead. Fluorescence resonance energy transfer between green fluorescent protein donors within the phage interior and acceptor-labeled DNA was used to confirm DNA packaging. Without ATP, no packaging was observed, and there was no evidence of substrate association with the prohead.
摘要
利用荧光相关光谱法在体外研究了噬菌体T4的DNA包装机制。通过监测伴随的DNA扩散性降低，测量了标记DNA从本体溶液到噬菌体内部的ATP依赖性转位动力学。发现单个噬菌体原头部可以依次包装多个短DNA片段（100个碱基对）。利用噬菌体内部绿色荧光蛋白供体与受体标记DNA之间的荧光共振能量转移来确认DNA包装。没有ATP时，未观察到包装现象，也没有证据表明底物与原头部结合。
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