title: A vector library for silencing central carbon metabolism genes with antisense RNAs in Escherichia coli.
abstract: We describe here the construction of a series of 71 vectors to silence central carbon metabolism genes in Escherichia coli. The vectors inducibly express antisense RNAs called paired-terminus antisense RNAs, which have a higher silencing efficacy than ordinary antisense RNAs. By measuring mRNA amounts, measuring activities of target proteins, or observing specific phenotypes, it was confirmed that all the vectors were able to silence the expression of target genes efficiently. Using this vector set, each of the central carbon metabolism genes was silenced individually, and the accumulation of metabolites was investigated. We were able to obtain accurate information on ways to increase the production of pyruvate, an industrially valuable compound, from the silencing results. Furthermore, the experimental results of pyruvate accumulation were compared to in silico predictions, and both sets of results were consistent. Compared to the gene disruption approach, the silencing approach has an advantage in that any E. coli strain can be used and multiple gene silencing is easily possible in any combination.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Sciences and Technology, Toyohira-ku, Sapporo, Japan.
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Nakashima
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ohno
authors.forename: Katsunori
authors.initials: K
authors.lastname: Yoshikawa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Tamura
mesh_terms: D002244:Carbon / Q000378:metabolism*; D003198:Computer Simulation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D015723:Gene Library; D020868:Gene Silencing; D005822:Genetic Vectors; D019289:Pyruvic Acid / Q000378:metabolism; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D002244:Carbon; D019289:Pyruvic Acid