Selection analyses of insertional mutants using subgenic-resolution arrays
Selection analyses of insertional mutants using subgenic-resolution arrays
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DOI：
10.1038/nbt1101-1060
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发表时间：
2001-11-01
影响因子：
46.9
通讯作者：
Church, GM
中科院分区：
文献类型：
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作者：
Badarinarayana, V;Estep, PW;Church, GM
我们描述了一种利用插入诱变和DNA微阵列对数量生长表型进行全基因组分析的方法。我们应用该方法评估了大肠杆菌基因域在特定生长条件下对适应性的贡献。一个转座子文库在卢里亚 - 贝尔塔尼（LB）培养基和葡萄糖基本培养基中进行竞争性生长筛选。将来自筛选文库的含转座子的基因组DNA片段与DNA微阵列上最初未筛选的转座子插入文库进行比较，以识别影响适应性的插入。发现在生长培养基中未提供的营养物质生物合成所涉及的基因在含有负选择插入的基因集中显著富集。这些数据还确定了几个未表征基因对适应性的贡献，包括假定的转录调节因子和酶。讨论了这种高分辨率阵列筛选在其他物种中的适用性。
We describe a method of genome-wide analysis of quantitative growth phenotypes using insertional mutagenesis and DNA microarrays. We applied the method to assess the fitness contributions of Escherichia coli gene domains under specific growth conditions. A transposon library was subjected to competitive growth selection in Luria-Bertani (LB) and in glucose minimal media. Transposon-containing genomic DNA fragments from the selected libraries were compared with the initial unselected transposon insertion library on DNA microarrays to identify insertions that affect fitness. Genes involved in the biosynthesis of nutrients not provided in the growth medium were found to be significantly enriched in the set of genes containing negatively selected insertions. The data also identify fitness contributions of several uncharacterized genes, including putative transcriptional regulators and enzymes. The applicability of this high-resolution array selection in other species is discussed.