Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3)

Amit Kumar Chaudhary, Je Won Park, Yeo Joon Yoon, Byung Gee Kim, Jae Kyung Sohng

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)285-293
Number of pages9
JournalBiotechnology letters
Volume35
Issue number2
DOIs
Publication statusPublished - 2013 Jan 1
Externally publishedYes

Fingerprint

Genetic Engineering
Escherichia coli
Aminoglycosides
Genes
Registries
Polyacrylamide Gel Electrophoresis
Nucleotides
Gene Expression
Polymerase Chain Reaction
2-deoxystreptamine

Keywords

  • Multi-monocistronic
  • Polycistronic
  • Pseudo-operon
  • Re-engineering of genetic circuit
  • Synthetic biology

ASJC Scopus subject areas

  • Biotechnology

Cite this

Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3). / Chaudhary, Amit Kumar; Park, Je Won; Yoon, Yeo Joon; Kim, Byung Gee; Sohng, Jae Kyung.

In: Biotechnology letters, Vol. 35, No. 2, 01.01.2013, p. 285-293.

Research output: Contribution to journalArticle

Chaudhary, Amit Kumar ; Park, Je Won ; Yoon, Yeo Joon ; Kim, Byung Gee ; Sohng, Jae Kyung. / Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3). In: Biotechnology letters. 2013 ; Vol. 35, No. 2. pp. 285-293.
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