Examination of Knock Out Mutants for Sensitivity to Phloroglucinol
Abstract
Progress relates to Deliverable # 1: Identification of the proteins involved in export of phloroglucinol and determination of whether overexpression of these proteins increases the concentration and yield of microbe-synthesized phloroglucinol. Using transcriptome analysis, candidate genes were identified that may be involved in the export of phloroglucinol from the inside of the Escherichia coli catalyst to the culture medium. Gene expression in non-phloroglucinol synthesizing E. coli W3110 serA(DE3)/pBCl.l46 was compared with gene expression in phloroglucinol synthesizing E. coli W3110 serA(DE3)/pJA3.131A. For both strains, cells were grown in 2-L fermentation vessels under identical conditions. Following transcriptome analysis, an extensive list of genes were identified whose expression changed substantially when phloroglucinol was synthesized by the cells. (Please see Progress Report 1 ). In order to gauge the importance of each gene product on cellular sensitivity to phloroglucinol, plans were laid to knock out the genes whose expression had increased in the presence of phloroglucinol. To determine which gene knock-outs should be prepared, genes that were upregulated at least two-fold were examined to determine if they were membrane proteins. From the list of upregulated genes, 41 genes were identified either as membrane proteins or putative membrane proteins.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 2011
- Accession Number
- ADA548949
Entities
People
- Karen M. Frost