Lynch Michael D, Warnecke Tanya, Gill Ryan T
Department of Chemical and Biological Engineering, University of Colorado, ECCH 111, Campus Box 424, Boulder, Colorado 80309, USA.
Nat Methods. 2007 Jan;4(1):87-93. doi: 10.1038/nmeth946. Epub 2006 Nov 12.
We report a genome-wide, multiscale approach to simultaneously measure the effect that the increased copy of each gene and/or operon has on a desired trait or phenotype. The method involves (i) growth selections on a mixture of several different plasmid-based genomic libraries of defined insert sizes or SCALEs, (ii) microarray studies of enriched plasmid DNA, and a (iii) mathematical multiscale analysis that precisely identifies the relevant genetic elements. This approach allows for identification of all single open reading frames and larger multigene fragments within a genomic library that alter the expression of a given phenotype. We have demonstrated this method in Escherichia coli by monitoring, in parallel, a population of >10(6) genomic library clones of different insert sizes, throughout continuous selections over a period of 100 generations.
我们报告了一种全基因组、多尺度方法，用于同时测量每个基因和/或操纵子的拷贝数增加对所需性状或表型的影响。该方法包括：（i）在几种不同的基于质粒的、具有特定插入片段大小或规模（SCALEs）的基因组文库混合物上进行生长筛选；（ii）对富集的质粒DNA进行微阵列研究；以及（iii）精确识别相关遗传元件的数学多尺度分析。这种方法能够识别基因组文库中所有改变给定表型表达的单个开放阅读框和更大的多基因片段。我们通过在100代的连续筛选过程中，并行监测超过10⁶个不同插入片段大小的基因组文库克隆群体，在大肠杆菌中证明了该方法。
