Genetic Manipulation Techniques in Streptomyces Species

Streptomyces are filamentous actinobacteria renowned for producing clinically important natural products. Genetic manipulation in these organisms underpins strain improvement, natural product discovery and synthetic biology applications. Early methods relied on protoplast transformation, where cell walls are enzymatically removed to allow uptake of plasmid DNA under osmotic control. Intergeneric conjugation from Escherichia coli to Streptomyces using specialised donor strains and integrative vectors harnessing phage-derived recombinases has become routine, offering robust chromosome integration. The advent of site-specific recombination systems, such as φC31 and pSAM2 integrases, enables marker-free gene insertions or deletions. Homologous recombination remains a mainstay for precise gene replacement, though efficiency can be hampered by low recombination rates. Recent innovations include CRISPR/Cas9-based genome editing and CRISPRi-mediated transcriptional silencing, which afford rapid and multiplexed target manipulation. Synthetic biology approaches have produced modular shuttle vectors and promoter libraries tailored for Streptomyces. Combined, these techniques have transformed genetic tractability, facilitating activation of cryptic biosynthetic gene clusters, metabolic pathway engineering and the discovery of novel bioactive compounds with global significance in medicine and biotechnology.

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Genetic Manipulation Techniques in Streptomyces Species publication trend

The graph below shows the total number of articles in genetic manipulation techniques in streptomyces species across all publications each year (not limited to Nature Index journals).

Technical terms

Protoplast: A bacterial cell from which the rigid cell wall has been removed, allowing direct DNA uptake.

Intergeneric conjugation: A process of plasmid transfer between different bacterial genera, typically from Escherichia coli to Streptomyces.

Site-specific recombination: Targeted insertion or excision of DNA segments at defined attP/attB sequences mediated by specialised integrases.

Integrative plasmid: A vector designed to integrate into the host chromosome via recombination, often carrying selectable markers.

Homologous recombination: Exchange of genetic material between DNA molecules with similar sequences, enabling precise gene replacement.

CRISPR/Cas9: An RNA-guided nuclease system adapted for targeted genome editing in diverse organisms, including Streptomyces.

References

1. Formation of Streptomyces protoplasts during cultivation in liquid media with lytic enzyme. Nova Biotechnologica et Chimica (2021).
2. Extremophile Metal Resistance: Plasmid-Encoded Functions in Streptomyces mirabilis. Applied and Environmental Microbiology (2022).
3. Molecular Characterization of Plasmids Harbored by Actinomycetes Isolated From the Great Salt Plains of Oklahoma Using PFGE and Next Generation Whole Genome Sequencing. Frontiers in Microbiology (2018).
4. Marker-Free Genome Engineering in Amycolatopsis Using the pSAM2 Site-Specific Recombination System. Microorganisms (2022).
5. Construct of Streptomyces Scabies Knock-Out Mutant of TXT Biosynthetic Gene txtB Via Intergeneric Conjugation. Biomedical Journal of Scientific & Technical Research (2018).
6. Construction of intergeneric conjugal transfer for molecular genetic studies of Streptomyces mobaraensis producing transglutaminase. African Journal of Biotechnology (2014).