Related Experiment Videos
Cloning vectors derived from Streptomyces niveus plasmid pSN2.
1Department of Genetics and Microbiology, Donnan Laboratories, University of Liverpool, UK.
Letters in Applied Microbiology
|November 1, 1990
Summary
New cloning vectors, pLG5 and pLG10, offer broad host range and high copy numbers for genetic engineering. These vectors facilitate cloning with selectable markers, enhancing Streptomyces research applications.
Area of Science:
- Molecular Biology
- Microbial Genetics
Background:
- Streptomyces plasmids are crucial tools for genetic manipulation.
- Development of stable, high-copy-number vectors is essential for efficient cloning and gene expression studies.
Purpose of the Study:
- To describe two novel, broad-host-range cloning vectors, pLG5 and pLG10, derived from Streptomyces niveus plasmid pSN2.
- To introduce a third vector, pLG505, with additional cloning capabilities.
Main Methods:
- Plasmid DNA isolation and characterization.
- Gene cloning and manipulation techniques.
- Transformation and selection of host bacteria.
Main Results:
- pLG5 (5.5 kb) and pLG10 (6.5 kb) vectors were successfully constructed and characterized.
- Both vectors contain thiostrepton resistance (TsrR) and lethal zygosis (Ltz+) markers.
- pLG505 (7.4 kb) was created by inserting the viomycin resistance (vph) gene into pLG5, providing insertion and replacement sites.
Conclusions:
- The described vectors (pLG5, pLG10, pLG505) are valuable tools for molecular cloning in a broad range of hosts.
- These vectors offer versatile options for gene insertion and selection, aiding genetic studies in Streptomyces and other bacteria.