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Pock forming plasmids from antibiotic-producing Streptomyces
Abstract:
Pock forming ability of fifteen plasmids isolated from antibiotic-producing Streptomyces was examined with polyethylene glycol mediated protoplast transformation. Four plasmids (pSF765, pSF689, pSF674 and pSF601) were found to have pock forming ability on Streptomyces lividans 66. Each putative transformants isolated from the center of pock regions harbored plasmids with same restriction enzyme cleavage sites as original plasmids. The plasmids from the transformants produced pock, which were morphologically identical to the original plasmids. Transformation frequency of S. lividans 66 with pSF689 from S. lividans 66 was at least 500-fold higher (5 x 10(5) transformants per 1 microgram DNA) than that with the plasmid isolated from the original strain (10(3) transformants per 1 microgram DNA). In case of pSF765 the transformation frequency was not changed (about 10(5) transformants per 1 microgram DNA) by replications in S. lividans 66. Expanded restriction enzyme maps for the four plasmids are reported.
Insights
Four plasmids from antibiotic-producing Streptomyces showed pock-forming ability in Streptomyces lividans 66 after polyethylene glycol-mediated protoplast transformation. This study details their characteristics and enhanced transformation frequencies.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Plasmids are crucial genetic elements in bacteria, often carrying genes for antibiotic resistance or production.
- Streptomyces species are well-known for producing a wide range of antibiotics, making their genetic manipulation important for biotechnology.
- Understanding plasmid behavior and transfer is key to harnessing antibiotic production capabilities.
Purpose of the Study:
- To investigate the pock-forming ability of plasmids isolated from antibiotic-producing Streptomyces.
- To characterize the genetic stability and transformation efficiency of these plasmids in a heterologous host, Streptomyces lividans 66.
- To construct expanded restriction enzyme maps for the identified pock-forming plasmids.
Main Methods:
- Isolation and characterization of fifteen plasmids from antibiotic-producing Streptomyces strains.
- Polyethylene glycol-mediated protoplast transformation of Streptomyces lividans 66.
- Analysis of pock formation, plasmid integrity (restriction enzyme mapping), and transformation frequency.
Main Results:
- Four plasmids (pSF765, pSF689, pSF674, and pSF601) exhibited pock-forming ability on S. lividans 66.
- Transformants harbored plasmids with identical restriction enzyme cleavage sites as the originals, indicating genetic stability.
- Transformation frequency of S. lividans 66 was significantly higher (up to 500-fold) using plasmids replicated in S. lividans 66 compared to those from the original strain.
- Transformation frequency for pSF765 remained stable after replication in S. lividans 66.
Conclusions:
- The identified plasmids possess the ability to form pocks and can be stably maintained and efficiently transformed in Streptomyces lividans 66.
- Replication in S. lividans 66 can enhance the transformation frequency of certain plasmids, like pSF689.
- Expanded restriction maps provide valuable tools for further genetic manipulation and study of these antibiotic-related plasmids.