通过绿色荧光蛋白（GFP）插入检测法测量插入序列IS1的转座频率。

Measurements of transposition frequency of insertion sequence IS1 by GFP hop-on assay.

作者信息

Saito Takashi, Chibazakura Taku, Takahashi Kiwamu, Yoshikawa Hirofumi, Sekine Yasuhiko

机构信息

Department of Life Science, College of Science, Rikkyo (St. Paul's) University, Toshima-ku, Tokyo, Japan.

出版信息

J Gen Appl Microbiol. 2010 Jun;56(3):187-92. doi: 10.2323/jgam.56.187.

DOI:10.2323/jgam.56.187

PMID:20647675

Abstract

Transposons play a significant role in the evolution of bacterial genomes. Quantifying frequency of transpositional events caused by a transposon will facilitate understanding its role. Here, we report successful measurement of the frequency of IS1 transposition using "GFP hop-on assay" in which transposition-dependent GFP expression is monitored by FACS. This assay allows easy assessment of IS transposition into the chromosomal DNA on a single-cell scale; this is an advantage over other conventional methods to measure transposition frequency.

摘要

转座子在细菌基因组的进化中起着重要作用。量化由转座子引起的转座事件的频率将有助于理解其作用。在此，我们报告了使用“GFP 插入测定法”成功测量 IS1 转座频率，其中通过流式细胞术监测转座依赖性 GFP 表达。该测定法允许在单细胞水平上轻松评估 IS 插入染色体 DNA 的情况；这是相对于其他测量转座频率的传统方法的一个优势。