Construction of a BioBrick™ compatible vector system for Rhodococcus

Plasmid. 2017 Mar:90:1-4. doi: 10.1016/j.plasmid.2017.01.004. Epub 2017 Jan 24.

Abstract

Throughout the past decade, the field of synthetic biology has grown rapidly. By using assembly platforms such as BioBricks™, scientists can quickly and easily build gene circuits or multi-step pathways. One limitation, however, is that most of these parts were designed and characterized with Escherichia coli as the target chassis. As a consequence, there exists a lack of standardized and well characterized or BioBrick™ compatible plasmid backbones that replicate in other potential non-model chassis organisms. The Gram-positive bacteria of the genus Rhodococcus represent an interesting chassis for biotechnological applications due to their tremendous metabolic capabilities. In this report we describe our progress toward developing a BioBrick™ compatible plasmid system for Rhodococcus. We demonstrate its utility for heterologous protein expression through flow cytometric analysis of the lac promoter in the oleaginous strain Rhodococcus opacus PD630.

Keywords: BioBrick™; Rhodococcus; Synthetic biology.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • DNA Restriction Enzymes / chemistry
  • DNA Restriction Enzymes / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genes, Reporter
  • Genetic Engineering / methods*
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Kanamycin / pharmacology
  • Lac Repressors / genetics*
  • Lac Repressors / metabolism
  • Plasmids / chemistry
  • Plasmids / metabolism*
  • Promoter Regions, Genetic / drug effects
  • Rhodococcus / drug effects
  • Rhodococcus / genetics*
  • Rhodococcus / metabolism

Substances

  • Anti-Bacterial Agents
  • Lac Repressors
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Kanamycin
  • DNA Restriction Enzymes