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使用亚基因分辨率阵列对插入突变体进行选择分析。

Selection analyses of insertional mutants using subgenic-resolution arrays.

作者信息

Badarinarayana V, Estep P W, Shendure J, Edwards J, Tavazoie S, Lam F, Church G M

机构信息

Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.

出版信息

Nat Biotechnol. 2001 Nov;19(11):1060-5. doi: 10.1038/nbt1101-1060.

Abstract

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.

摘要

我们描述了一种使用插入诱变和DNA微阵列对定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特定生长条件下大肠杆菌基因结构域对适应性的贡献。一个转座子文库在Luria-Bertani(LB)培养基和葡萄糖基本培养基中进行竞争性生长选择。将来自选定文库的含转座子的基因组DNA片段与初始未选择的转座子插入文库在DNA微阵列上进行比较,以鉴定影响适应性的插入。发现在含有负选择插入的基因集中,参与生长培养基中未提供的营养物质生物合成的基因显著富集。数据还确定了几个未表征基因的适应性贡献,包括假定的转录调节因子和酶。讨论了这种高分辨率阵列选择在其他物种中的适用性。

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