Protocol to develop a synthetic biology toolkit for the non-model bacterium R. palustris

Mark Kathol; Cheryl Immethun; Rajib Saha

doi:10.1016/j.xpro.2023.102158

Protocol to develop a synthetic biology toolkit for the non-model bacterium R. palustris

STAR Protoc. 2023 Apr 26;4(2):102158. doi: 10.1016/j.xpro.2023.102158. Online ahead of print.

Authors

Mark Kathol¹, Cheryl Immethun², Rajib Saha³

Affiliations

¹ Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA. Electronic address: mkathol2@huskers.unl.edu.
² Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
³ Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA. Electronic address: rsaha2@huskers.unl.edu.

Abstract

Numerous biology tools are developed to work for model organisms, which, however, do not work effectively in non-model organisms. Here, we present a protocol for developing a synthetic biology toolkit for Rhodopseudomonas palustris CGA009, a non-model bacterium with unique metabolic properties. We describe steps for introducing and characterizing biological devices in non-model bacteria, such as the utilization of fluorescence markers and RT-qPCR. This protocol may also be applicable for other non-model organisms. For complete details on the use and execution of this protocol, please refer to Immethun et al..¹.

Keywords: Biotechnology and Bioengineering; Gene Expression; Genetics; Metabolism; Microbiology; Molecular Biology; Molecular/Chemical Probes.