Isolation and characterization of Pseudomonas putida R-prime plasmids
Journal of General Microbiology
|March 1, 1987
Summary
Researchers constructed R-prime plasmids using enhanced chromosome mobilizing (ECM) plasmids from Pseudomonas putida. Different mechanisms may be involved in R-prime formation and chromosome mobilization by these plasmids.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Enhanced chromosome mobilizing (ECM) plasmids, derived from the wide host range plasmid R68, are tools for genetic manipulation.
- R-prime plasmids are essential for transferring chromosomal DNA between bacterial strains, facilitating genetic studies.
Purpose of the Study:
- To construct R-prime plasmids carrying specific segments of the Pseudomonas putida PPN chromosome.
- To investigate the mechanisms of R-prime formation and chromosome mobilization using ECM plasmids in Pseudomonas aeruginosa.
Main Methods:
- Utilized enhanced chromosome mobilizing (ECM) plasmids derived from R68.
- Constructed R-prime plasmids containing up to two map minutes of the Pseudomonas putida PPN chromosome.
- Employed Pseudomonas aeruginosa PAO as the recipient strain for R-prime construction.
Main Results:
- Successfully constructed R-prime plasmids carrying Pseudomonas putida chromosomal DNA.
- Observed that for ECM plasmid pMO61, R-prime formation ability did not correlate with intrastrain chromosome mobilization.
- Physical analysis revealed a 3.5 kb chromosomal DNA insertion within the IS21 sequences of a constructed R-prime.
Conclusions:
- The study demonstrates the successful construction of R-prime plasmids for Pseudomonas genetic studies.
- Findings suggest distinct mechanisms govern R-prime formation and chromosome mobilization mediated by ECM plasmids.
- The insertion of chromosomal DNA within the IS21 sequences is a key feature of the generated R-primes.
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