Developing a Synthetic Biology Toolkit for Comamonas testosteroni, an Emerging Cellular Chassis for Bioremediation

Qiang Tang, Ting Lu, Shuang Jiang Liu

Research output: Contribution to journalArticle

Abstract

Synthetic biology is rapidly evolving into a new phase that emphasizes real-world applications such as environmental remediation. Recently, Comamonas testosteroni has become a promising chassis for bioremediation due to its natural pollutant-degrading capacity; however, its application is hindered by the lack of fundamental gene expression tools. Here, we present a synthetic biology toolkit that enables rapid creation of functional gene circuits in C. testosteroni. We first built a shuttle system that allows efficient circuit construction in E. coli and necessary phenotypic testing in C. testosteroni. Then, we tested a set of wildtype inducible promoters, and further used a hybrid strategy to create engineered promoters to expand expression strength and dynamics. Additionally, we tested the T7 RNA Polymerase-PT7 promoter system and reduced its leaky expression through promoter mutation for gene expression. By coupling random library construction with FACS screening, we further developed a synthetic T7 promoter library to confer a wider range of expression strength and dynamic characteristics. This study provides a set of valuable tools to engineer gene circuits in C. testosteroni, facilitating the establishment of the organism as a useful microbial chassis for bioremediation purposes.

Original languageEnglish (US)
Pages (from-to)1753-1762
Number of pages10
JournalACS synthetic biology
Volume7
Issue number7
DOIs
StatePublished - Jul 20 2018

Fingerprint

Comamonas testosteroni
Synthetic Biology
Environmental Biodegradation
Bioremediation
Chassis
Gene expression
Networks (circuits)
Gene Regulatory Networks
Genes
Libraries
RNA
Gene Expression
Escherichia coli
Screening
Engineers
Testing
Mutation

Keywords

  • Comamonas testosteroni
  • P promoter library
  • T7 RNA polymerase-P system
  • inducible promoter
  • shuttle vehicle
  • synthetic biology toolkit

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Developing a Synthetic Biology Toolkit for Comamonas testosteroni, an Emerging Cellular Chassis for Bioremediation. / Tang, Qiang; Lu, Ting; Liu, Shuang Jiang.

In: ACS synthetic biology, Vol. 7, No. 7, 20.07.2018, p. 1753-1762.

Research output: Contribution to journalArticle

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