Baldwin David N, Salama Nina R
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Methods Enzymol. 2007;421:90-110. doi: 10.1016/S0076-6879(06)21010-3.
The rapid expanse of microbial genome databases provides incentive and opportunity to study organismal behavior at the whole-genome level. While many newly sequenced genes are assigned names based on homology to previously characterized genes, many putative open reading frames remain to be annotated. The use of microarrays enables functional characterization of the entire genome with respect to genes important for different growth conditions including nutrient deprivation, stress responses, and virulence. The methods described here combine advancements in the identification of genomic sequences flanking insertional mutants with microarray methodology. The combination of these methods facilitates tracking large numbers of mutants for phenotypic studies. This improves both the efficiency of genome-saturating library screens and contributes to the functional annotation of unknown genes.
微生物基因组数据库的迅速扩展为在全基因组水平上研究生物体行为提供了动力和机会。虽然许多新测序的基因是根据与先前已表征基因的同源性来命名的,但仍有许多假定的开放阅读框有待注释。微阵列的使用能够针对对不同生长条件(包括营养剥夺、应激反应和毒力)重要的基因对整个基因组进行功能表征。本文所述方法将插入突变体侧翼基因组序列鉴定方面的进展与微阵列方法相结合。这些方法的结合有助于追踪大量用于表型研究的突变体。这提高了基因组饱和文库筛选的效率,并有助于对未知基因进行功能注释。