Artificial RNA aptamer production by the marine bacterium Rhodovulum sulfidophilum : Improvement of the aptamer yield using a mutated transcriptional promoter(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
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概要
- 論文の詳細を見る
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 200ng and the total RNA aptamer (both extra- and intracellular form) of about 20μg from 1L culture were accomplished by constitutive expression of the gene.
著者
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Tanaka T
Bio-organic Chemistry Laboratory Institute For Fundamental Research Suntory Research Center
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Tanaka Takaaki
Graduate School Of Science And Technology Niigata University
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Kikuchi Y
Div. Of Life Sci. And Biotechnology Dep. Of Ecological Engineering Toyohashi Univ. Of Technol.
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Kikuchi Yo
Division Of Bioscience And Biotechnology Department Of Ecological Engeneering Toyohashi University O
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Umekage So
Division Of Life Science And Biotechnology Department Of Ecological Engineering Toyohashi University
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Tanaka T
Division Of Life Science And Biotechnology Department Of Ecological Engineering Toyohashi University
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Kikuchi Y
Division Of Bioscience And Biotechnology Department Of Ecological Engineering Toyohashi University O
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Suzuki Hiromichi
Division Of Life Science And Biotechnology Department Of Ecological Engineering Toyohashi University
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Tanaka Terumichi
Division Of Bioscience And Biotechnology Department Of Ecological Engeneering Toyohashi University O
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Umekage So
Division Of Life Science And Biotechnology Department Of Ecological Engineering Toyohashi University
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Tanaka Takaaki
Department Of Materials Science And Technology Faculty Of Engineering Niigata University
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Suzuki Hiromichi
Division Of Bioscience And Biotechnology Department Of Environmental And Life Sciences Toyohashi Uni
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Umekage So
Division Of Bioscience And Biotechnology Department Of Environmental And Life Sciences Toyohashi Uni
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Tanaka Terumichi
Graduate School of Science and Technology, Niigata University
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