pSHDY: A New Tool for Genetic Engineering of Cyanobacteria
Abstract
Genetic engineering of cyanobacteria is currently limited to genomic integration via homologous recombination and RSF1010-based conjugative vector systems. Here, we introduce a rationally designed conjugative vector with two BioBrick-based cloning sites which enables facilitated and modular cloning. This streamlined vector is suitable for a variety of synthetic biology applications, such as expression of multiple enzymes from metabolic pathways for the production of biofuels or secondary metabolites, or screening of modular parts such as promoters, further facilitating applications to improve crop plants using synthetic biology. Finally, we present a general approach to cloning of constructs, as well as detailed protocols for conjugation and culturing of strains carrying said constructs.
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Shuttle plasmids can replicate in multiple host organisms, usually Escherichia coli and the target cyanobacterial strain, which simplifies plasmid construction and amplification in E. coli prior to transfer into cyanobacteria.
“Shuttle plasmids can replicate in multiple host organisms, usually Escherichia coli and the target cyanobacterial strain, which simplifies plasmid construction and amplification in E. coli prior to transfer into cyanobacteria [88,90].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [88]
The selection between plasmid-based expression and chromosomal integration depends on several factors, including the required stability of gene expression, the size of the introduced construct, and the specific physiological and genetic characteristics of the cyanobacterial strain.
“The selection between these methods depends on several factors, including the required stability of gene expression, the size of the introduced construct, and the specific physiological and genetic characteristics of the cyanobacterial strain [88].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [88]
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“The selection between these methods depends on several factors, including the required stability of gene expression, the size of the introduced construct, and the specific physiological and genetic characteristics of the cyanobacterial strain [88].”
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- Authors
- Behle, A., & Axmann, I. M.
- Affiliations
- Heinrich Heine University Düsseldorf (DE)
- Corresponding author
- Axmann, Ilka M.
- Venue
- Methods in Molecular Biology
- Publisher
- Springer US
- Published
- 2022
- Volume
- 2379
- Pages
- 67-79
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- OpenAlex
- W4213273407
- Cited by
- 10
- References
- 30
- Citations by year
- 2026: 5 · 2025: 1 · 2023: 3 · 2022: 1
- FWCI
- 2.10
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Keywords
- cyanobacteria, genetic engineering, homologous recombination, RSF1010, synthetic biology, biofuel production, secondary metabolite production, promoter screening, crop improvement, conjugation
- MeSH terms
- Cyanobacteria, Genetic Engineering, Genetic Vectors, Plasmids, Synthetic Biology, Metabolic Engineering
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