Expression and characterization of the complete cyanophage genome PP in the heterologous host Synechococcus elongatus PCC 7942

Guorui Li; Jia Feng; Xiaofei Zhu; Yujie Chai; Tao Sun; Jianlan Jiang

doi:10.1016/j.ijbiomac.2025.140242

Expression and characterization of the complete cyanophage genome PP in the heterologous host Synechococcus elongatus PCC 7942

Int J Biol Macromol. 2025 Jan 23:301:140242. doi: 10.1016/j.ijbiomac.2025.140242. Online ahead of print.

Authors

Guorui Li¹, Jia Feng², Xiaofei Zhu³, Yujie Chai⁴, Tao Sun⁵, Jianlan Jiang⁶

Affiliations

¹ School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, PR China; Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China.
² School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, PR China; Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, 300072, PR China.
³ School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, PR China; State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, 300072, PR China.
⁴ School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, PR China.
⁵ School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, PR China; Center for Biosafety Research and Strategy, Tianjin University, Tianjin 300072, PR China; State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, 300072, PR China. Electronic address: tsun@tju.edu.cn.
⁶ School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, PR China; Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, PR China; Frontier Science Center for Synthetic Biology (Ministry of Education), Tianjin University, Tianjin 300072, PR China; State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin, 300072, PR China. Electronic address: jljiang@tju.edu.cn.

PMID: 39863235
DOI: 10.1016/j.ijbiomac.2025.140242

Abstract

In this study, we successfully integrated the full-length genome of the cyanophage PP into the non-host cyanobacterium Synechococcus elongatus PCC 7942, facilitated by conjugation via Escherichia coli. To address the challenge posed by the toxic open reading frames (ORFs) of PP in E. coli, we first identified and characterized three toxic ORFs. The PP genome was subsequently rearranged, and the expression of these toxic ORFs was controlled using a tandem-induction switch system. The full-length PP genome was then successfully integrated into the genome of S. elongatus PCC 7942. Interestingly, the integration of the PP genome led to a reduction in photosynthesis and carbon fixation in S. elongatus PCC 7942, resembling the effects typically associated with cyanophage infection. Transcriptomic analysis showed that 32 of the 41 ORFs in the PP genome were actively transcribed in S. elongatus PCC 7942, significantly affecting energy metabolism and carbon fixation pathways. These effects were further confirmed by metabolomic analysis.

Keywords: Cyanobacteria; Cyanophage; Genome design and assembly; Inducible switches; Transcriptomics.