title: A screening system for artificial small RNAs that inhibit the growth of Escherichia coli.
abstract: We have developed a screening system for artificial small RNAs (sRNAs) that inhibit the growth of Escherichia coli. In this system, we used a plasmid library to express artificial sRNAs (approximately 200 bases long) containing 60 bases of random nucleotide sequence. The induced expression of the known rydB sRNA in the system reduced the amount of its possible target mRNA, rpoS, supporting the reliability of the method. To isolate clones of sRNAs that inhibited the growth of E. coli, we used two successive screening steps: (i) colony size selection on plates and (ii) monitoring E. coli growth in a 96-well plate format. As a result, 83 artificial sRNAs were identified that showed a range of inhibitory effects on bacterial growth. We also introduced nucleotide replacements into one of the highly inhibitory sRNA clones, H12, which partially abolished the inhibition of bacterial growth, suggesting that bacterial growth was inhibited in a sequence-specific manner.
authors.affiliation: Institute for Advanced Biosciences, Keio University, Tsuruoka 997-0017, Japan.
authors.forename: Mizuki
authors.initials: M
authors.lastname: Komasa
authors.forename: Kosuke
authors.initials: K
authors.lastname: Fujishima
authors.forename: Kiriko
authors.initials: K
authors.lastname: Hiraoka
authors.forename: Atsuko
authors.initials: A
authors.lastname: Shinhara
authors.forename: Baek-Seok
authors.initials: BS
authors.lastname: Lee
authors.forename: Masaru
authors.initials: M
authors.lastname: Tomita
authors.forename: Akio
authors.initials: A
authors.lastname: Kanai
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development*; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D012329:RNA, Bacterial; D012333:RNA, Messenger / Q000037:antagonists & inhibitors*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D012329:RNA, Bacterial; D012333:RNA, Messenger; D034741:RNA, Small Interfering