Your browser doesn't support javascript.
loading
Optimizing dsRNA engineering strategies and production in E. coli HT115 (DE3).
da Rosa, Juliana; Viana, Américo José Carvalho; Ferreira, Fernando Rafael Alves; Koltun, Alessandra; Mertz-Henning, Liliane Marcia; Marin, Silvana Regina Rockenbach; Rech, Elibio Leopoldo; Nepomuceno, Alexandre Lima.
Afiliação
  • da Rosa J; Department of General Biology, Londrina State University, Celso Garcia Cid Road, PR 445, km 380, University Campus, 86057-970 Londrina, PR, Brazil.
  • Viana AJC; Embrapa Soja, Carlos João Strass Highway, Acess Orlando Amaral, District of Warta, 86085-981 Londrina, PR, Brazil.
  • Ferreira FRA; Embrapa Soja, Carlos João Strass Highway, Acess Orlando Amaral, District of Warta, 86085-981 Londrina, PR, Brazil.
  • Koltun A; Arthur Bernardes Foundation, Headquarters Building, no number - University Campus, 36570-900 Viçosa, MG, Brazil.
  • Mertz-Henning LM; Embrapa Soja, Carlos João Strass Highway, Acess Orlando Amaral, District of Warta, 86085-981 Londrina, PR, Brazil.
  • Marin SRR; Arthur Bernardes Foundation, Headquarters Building, no number - University Campus, 36570-900 Viçosa, MG, Brazil.
  • Rech EL; Embrapa Soja, Carlos João Strass Highway, Acess Orlando Amaral, District of Warta, 86085-981 Londrina, PR, Brazil.
  • Nepomuceno AL; Embrapa Soja, Carlos João Strass Highway, Acess Orlando Amaral, District of Warta, 86085-981 Londrina, PR, Brazil.
J Ind Microbiol Biotechnol ; 512024 Jan 09.
Article em En | MEDLINE | ID: mdl-39152090
ABSTRACT
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%.
Assuntos
Palavras-chave
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA de Cadeia Dupla / Meios de Cultura / Escherichia coli / Fermentação Idioma: En Revista: J Ind Microbiol Biotechnol Assunto da revista: BIOTECNOLOGIA / MICROBIOLOGIA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Alemanha
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA de Cadeia Dupla / Meios de Cultura / Escherichia coli / Fermentação Idioma: En Revista: J Ind Microbiol Biotechnol Assunto da revista: BIOTECNOLOGIA / MICROBIOLOGIA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Alemanha