Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3)
- 주제(키워드) Multi-monocistronic , Polycistronic , Pseudo-operon , Re-engineering of genetic circuit , Synthetic biology
- 등재 SCIE, SCOPUS
- 발행기관 Kluwer Academic Publishers
- 발행년도 2013
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000094602
- 본문언어 영어
- Published As http://dx.doi.org/10.1007/s10529-012-1077-2
초록/요약
Various approaches for monocistronic constructions of genetic circuits have been designed for metabolite production but there has been no attempt to apply such methodology for aminoglycosides biosynthesis. Here, a simple and commercially available bio-part, despite the current trend focusing on the standardized BioBricks bio-parts available in the registry, is used. A 181-bp nucleotide fragment was designed for the efficient construction of an expression vector for monocistronic assembly of genes. Furthermore, a single vector with multi-monocistronic assembled genes for 2-deoxystreptamine (2-DOS) synthesis was constructed for production in engineered Escherichia coli. The working efficiency of model vector was concluded by reporter assay whereas the expressions of biosynthesis genes were confirmed by RT-PCR and SDS-PAGE. Production of 2-DOS was confirmed by TLC, LC-ELSD, and ESI-MS/MS. © 2012 Springer Science+Business Media Dordrecht.
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