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| 12. |  | SIMÕES-ARAÚJO, J. L. de; SILVA-JÚNIOR, Z.; MENESES, C. H. S. G.; VIDAL, M. S. Expression of VuNIP1 (NOD26-like) in cowpea nodules under water stress. In: ANNUAL MEETING OF SBBq, 37., CONGRESS OF THE PABMB, 11., 2008, Águas de Lindóia. Abstracts... Águas de Lindóia: SBBq, 2008. 1 p. Parceria: UFRJ.| Biblioteca(s): Embrapa Agrobiologia. |
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| Registros recuperados : 88 | |
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 | Acesso ao texto completo restrito à biblioteca da Embrapa Soja. Para informações adicionais entre em contato com valeria.cardoso@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Recursos Genéticos e Biotecnologia; Embrapa Soja. |
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Data corrente: |
14/10/2024 |
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Data da última atualização: |
14/10/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
A - 2 |
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Autoria: |
ROSA, J. da; VIANA, A. J. C.; FERREIRA, F. R. A.; KOLTUN, A.; MERTZ-HENNING, L. M.; MARIN, S. R. R.; RECH FILHO, E. L.; NEPOMUCENO, A. L. |
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Afiliação: |
JULIANA DA ROSA, LONDRINA STATE UNIVERSITY; AMÉRICO JOSÉ CARVALHO VIANA; FERNANDO RAFAEL ALVES FERREIRA; ALESSANDRA KOLTUN; LILIANE MARCIA MERTZ HENNING, CNPSO; SILVANA REGINA ROCKENBACH MARIN, CNPSO; ELIBIO LEOPOLDO RECH FILHO, CENARGEN; ALEXANDRE LIMA NEPOMUCENO, CNPSO. |
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Título: |
Optimizing dsRNA engineering strategies and production in E. coli HT115 (DE3). |
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Ano de publicação: |
2024 |
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Fonte/Imprenta: |
Journal of Industrial Microbiology and Biotechnology, v. 51, kuae028, 2024. |
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DOI: |
https://doi.org/10.1093/jimb/kuae028 |
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Idioma: |
Inglês |
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Notas: |
Na publicação: Elibio Leopoldo Rech. |
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Conteúdo: |
Producing double-stranded RNA (dsRNA) represents a bottleneck for the adoption of RNA interference technology in agriculture, and the main hurdles are related to increases in dsRNA yield, production efficiency, and purity. Therefore, this study aimed to optimize dsRNA production in E. coli HT115 (DE3) using an in vivo system. To this end, we designed a new vector, pCloneVR_2, which resulted in the efficient production of dsRNA in E. coli HT115 (DE3). We performed optimizations in the culture medium and expression inducer in the fermentation of E. coli HT115 (DE3) for the production of dsRNA. Notably, the variable that had the greatest effect on dsRNA yield was cultivation in TB medium, which resulted in a 118% increase in yield. Furthermore, lactose induction (6 g/L) yielded 10 times more than IPTG. Additionally, our optimized up-scaled protocol of the TRIzol™ extraction method was efficient for obtaining high-quality and pure dsRNA. Finally, our optimized protocol achieved an average yield of 53.3 µg/mL after the production and purification of different dsRNAs, reducing production costs by 72%. |
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Palavras-Chave: |
Bacterial dsRNA production; Lactose induction; RNAi. |
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Thesaurus NAL: |
Double-stranded RNA. |
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Categoria do assunto: |
--
X Pesquisa, Tecnologia e Engenharia |
|
Marc: |
LEADER 01991naa a2200277 a 4500
001 2168059
005 2024-10-14
008 2024 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1093/jimb/kuae028$2DOI
100 1 $aROSA, J. da
245 $aOptimizing dsRNA engineering strategies and production in E. coli HT115 (DE3).$h[electronic resource]
260 $c2024
500 $aNa publicação: Elibio Leopoldo Rech.
520 $aProducing double-stranded RNA (dsRNA) represents a bottleneck for the adoption of RNA interference technology in agriculture, and the main hurdles are related to increases in dsRNA yield, production efficiency, and purity. Therefore, this study aimed to optimize dsRNA production in E. coli HT115 (DE3) using an in vivo system. To this end, we designed a new vector, pCloneVR_2, which resulted in the efficient production of dsRNA in E. coli HT115 (DE3). We performed optimizations in the culture medium and expression inducer in the fermentation of E. coli HT115 (DE3) for the production of dsRNA. Notably, the variable that had the greatest effect on dsRNA yield was cultivation in TB medium, which resulted in a 118% increase in yield. Furthermore, lactose induction (6 g/L) yielded 10 times more than IPTG. Additionally, our optimized up-scaled protocol of the TRIzol™ extraction method was efficient for obtaining high-quality and pure dsRNA. Finally, our optimized protocol achieved an average yield of 53.3 µg/mL after the production and purification of different dsRNAs, reducing production costs by 72%.
650 $aDouble-stranded RNA
653 $aBacterial dsRNA production
653 $aLactose induction
653 $aRNAi
700 1 $aVIANA, A. J. C.
700 1 $aFERREIRA, F. R. A.
700 1 $aKOLTUN, A.
700 1 $aMERTZ-HENNING, L. M.
700 1 $aMARIN, S. R. R.
700 1 $aRECH FILHO, E. L.
700 1 $aNEPOMUCENO, A. L.
773 $tJournal of Industrial Microbiology and Biotechnology$gv. 51, kuae028, 2024.
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