Dynamics of genomic-library enrichment and identification of solvent tolerance genes for Clostridium acetobutylicum.

ABSTRACT: A Clostridium acetobutylicum ATCC 824 genomic library was constructed using randomly sheared DNA. Library inserts conferring increased tolerance to 1-butanol were isolated using two protocols. Protocol I utilized a single round of butanol challenges in batch culture, while protocol II, which gave clearly superior outcomes, was based on the serial transfer of stationary-phase cultures into progressively higher butanol concentrations. DNA microarray analysis made a high-resolution assessment of the dynamic process of library enrichment possible for the first time. Protocol I yielded a library insert containing the entire coding region of the gene CAC0003 (which codes for a protein of unknown function) but also several DNA fragments containing promoter regions. Protocol II enabled the success

SUBMITTER: Borden JR 

PROVIDER: S-EPMC1892849 | biostudies-literature | 2007 May

REPOSITORIES: biostudies-literature