Artificial RNA aptamer production by the marine bacterium Rhodovulum sulfidophilum

Improvement of the aptamer yield using a mutated transcriptional promoter

Hiromichi Suzuki, So Umekage, Terumichi Tanaka, Yo Kikuchi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Noncoding small RNAs and artificial RNA aptamers are now expected to be potential candidates for RNA therapeutic agents. We previously proposed a unique method for economical production of these RNAs using the marine phototrophic bacterium Rhodovulum sulfidophilum. This bacterium does not produce any ribonucleases but does produce extracellular nucleic acids in the culture medium in nature. Using this bacterium and an engineered plasmid containing the rrn promoter for the RNA expression, we developed a method for production of the streptavidin RNA aptamer in the culture medium. However, the yield of this RNA product in the culture medium by this method was not enough for practical use. In the present paper, we improved the yield of this product by modification of the - 35 region of the rrn promoter so as to escape from the Fis protein control and the use of a new vector plasmid. Using this system, the extracellular RNA aptamer of approximately 200. ng and the total RNA aptamer (both extra- and intracellular form) of about 20 μg from 1. L culture were accomplished by constitutive expression of the gene.

Original languageEnglish
Pages (from-to)458-461
Number of pages4
JournalJournal of Bioscience and Bioengineering
Volume112
Issue number5
DOIs
Publication statusPublished - 2011 Nov
Externally publishedYes

Fingerprint

Rhodovulum
Nucleotide Aptamers
RNA
Bacteria
Culture Media
Plasmids
Small Untranslated RNA
Streptavidin
Ribonucleases
Cell culture
Genetic Promoter Regions
Nucleic Acids
Genes
Gene Expression
Nucleic acids
Proteins

Keywords

  • Functional RNA
  • Phototrophic bacterium
  • Promoter
  • RNA aptamer
  • Transcription

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Artificial RNA aptamer production by the marine bacterium Rhodovulum sulfidophilum : Improvement of the aptamer yield using a mutated transcriptional promoter. / Suzuki, Hiromichi; Umekage, So; Tanaka, Terumichi; Kikuchi, Yo.

In: Journal of Bioscience and Bioengineering, Vol. 112, No. 5, 11.2011, p. 458-461.

Research output: Contribution to journalArticle

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