Construction and characterization of a copy number-inducible fosmid library of Xanthomonas oryzae pathovar oryzae MAFF311018
Construction and characterization of a copy number-inducible fosmid library of Xanthomonas oryzae pathovar oryzae MAFF311018
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DOI：
10.1016/j.gene.2014.05.035
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发表时间：
2014-08-01
期刊：
影响因子：
3.5
通讯作者：
Ohsato, Shuichi
中科院分区：
文献类型：
--
作者：
Ichida, Hiroyuki;Sun, Xiaoying;Ohsato, Shuichi
构建了水稻白叶枯病病原菌--水稻白叶枯病菌MAFF311018(T7174)的融合体文库，并对其进行了鉴定。Fosmid文库插入的平均大小为34kb，967个克隆在其测序的基因组上唯一定位。Xoo MAFF311018全基因组被末端测序的克隆覆盖，至少有5kb的重叠。Fosid载体既包含单拷贝的大肠杆菌育性因子起始点，增强了融合体的稳定性，又含有多拷贝的INCPα起始点，允许在L-阿拉伯糖诱导下扩增拷贝数。对随机挑选的12个Fosid文库克隆进行实时定量聚合酶链式反应，结果表明，诱导5小时后，Fosid拷贝数增加了8~58倍。该文库为Xoo及其近缘物种的互补实验和系统功能研究提供了新的资源。(C)爱思唯尔出版的2014年。
A fosmid library of Xanthomonas oryzae pathovar oryzae MAFF311018 (T7174), the causative agent of bacterial blight on rice, was constructed and characterized. The average fosmid library insert size was >34 kb, and 967 clones were uniquely positioned on its sequenced genome. The entire Xoo MAFF311018 genome was covered by end-sequenced clones with at least 5 kb of overlap. The fosmid vector contains both the single-copy Escherichia coli fertility factor origin, which enhances fosmid stability, and the multi-copy IncP alpha origin, allowing amplification of copy number upon induction with L-arabinose. Real-time quantitative PCR on 12 randomly picked fosmid library clones determined that fosmid copy number increased 8- to 58-fold after 5 hour induction. This library provides a new resource for complementation experiments and systematic functional studies in Xoo and related species. (C) 2014 Published by Elsevier B.V.