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.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Komasa M,Fujishima K,Hiraoka K,Shinhara A,Lee BS,Tomita M,Kanai A

doi

10.1093/jb/mvr055

subject

Has Abstract

pub_date

2011-09-01 00:00:00

pages

289-94

issue

3

eissn

0021-924X

issn

1756-2651

pii

mvr055

journal_volume

150

pub_type

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