A screening system for artificial small RNAs that inhibit the growth of Escherichia coli

Mizuki Komasa, Kosuke Fujishima, Kiriko Hiraoka, Atsuko Shinhara, Baek Seok Lee, Masaru Tomita, Akio Kanai

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)289-294
Number of pages6
JournalJournal of Biochemistry
Volume150
Issue number3
DOIs
Publication statusPublished - 2011 Sep

Fingerprint

Escherichia coli
Screening
RNA
Growth
Nucleotides
Clone Cells
Plasmids
Messenger RNA
Monitoring

Keywords

  • bacterial growth
  • biotechnology
  • Escherichia coli
  • screening system
  • small RNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

A screening system for artificial small RNAs that inhibit the growth of Escherichia coli. / Komasa, Mizuki; Fujishima, Kosuke; Hiraoka, Kiriko; Shinhara, Atsuko; Lee, Baek Seok; Tomita, Masaru; Kanai, Akio.

In: Journal of Biochemistry, Vol. 150, No. 3, 09.2011, p. 289-294.

Research output: Contribution to journalArticle

Komasa, Mizuki ; Fujishima, Kosuke ; Hiraoka, Kiriko ; Shinhara, Atsuko ; Lee, Baek Seok ; Tomita, Masaru ; Kanai, Akio. / A screening system for artificial small RNAs that inhibit the growth of Escherichia coli. In: Journal of Biochemistry. 2011 ; Vol. 150, No. 3. pp. 289-294.
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