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Intraplasmid recombination in Streptomyces lividans 66
C W Chen1, J F Tsai, S E Chuang
1Institute of Microbiology and Immunology, National Yangming Medical College, Taipei, Taiwan, Republic of China.
Molecular & General Genetics : MGG
|August 1, 1987
Summary
A new shuttle plasmid showed instability in Streptomyces lividans due to mel repeat recombination, forming a smaller plasmid. It also formed oligomers in E. coli but not in S. lividans.
Area of Science:
- Molecular Biology
- Microbial Genetics
- Plasmid Biology
Background:
- Shuttle plasmids are essential tools for genetic manipulation across different bacterial species.
- Understanding plasmid stability is crucial for efficient gene expression and cloning in hosts like Escherichia coli and Streptomyces lividans.
Purpose of the Study:
- To construct and characterize an Escherichia coli-Streptomyces shuttle plasmid, pIF132.
- To investigate the structural stability and replication behavior of pIF132 in both E. coli and S. lividans.
Main Methods:
- Construction of the pIF132 shuttle plasmid containing two direct mel repeats.
- Assessment of plasmid replication and structural stability in E. coli (Rec+ and recA strains).
- Evaluation of plasmid replication and structural stability in Streptomyces lividans.
Main Results:
- The shuttle plasmid pIF132 exhibited structural instability in S. lividans, with recombination between mel repeats yielding a smaller plasmid, pIF138.
- pIF132 demonstrated stable replication in E. coli, irrespective of Rec+ or recA status.
- Extensive oligomerization of pIF132 was observed in E. coli, a phenomenon not seen in S. lividans.
Conclusions:
- The direct mel repeats in pIF132 are prone to recombination in S. lividans, leading to plasmid instability.
- Differential oligomerization patterns exist between E. coli and S. lividans for the pIF132 plasmid.
- Plasmid stability mechanisms vary significantly between E. coli and S. lividans, impacting shuttle vector design.