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Replication origins of single-stranded-DNA plasmid pUB110
1Laboratoire de Génétique Microbienne, INRA-Domaine de Vilvert, Jouy en Josas, France.
Journal of Bacteriology
|June 1, 1989
Summary
The study characterizes plasmid pUB110 replication origins. Findings suggest rolling circle replication for the plus origin and RNA polymerase involvement in minus origin function, which is functional across multiple hosts.
Area of Science:
- Molecular Biology
- Microbiology
- Genetics
Background:
- Plasmid replication is crucial for bacterial genetics and biotechnology.
- Understanding replication origins (ori) is key to controlling plasmid maintenance and function.
- Plasmid pUB110 is a well-characterized Gram-positive bacterial plasmid.
Purpose of the Study:
- To characterize the two replication origins of plasmid pUB110.
- To determine the minimal functional sequences for each origin.
- To elucidate the replication mechanism and host range of pUB110.
Main Methods:
- DNA sequencing and manipulation to define origin boundaries.
- In vivo replication assays.
- Sensitivity testing with antibiotics like rifampin.
Main Results:
- The plus origin initiation site was mapped to an 8-bp sequence.
- Evidence suggests pUB110 utilizes rolling circle replication.
- The minus origin's minimal sequence was determined, showing orientation specificity.
- Minus origin activity is sensitive to rifampin, indicating RNA polymerase involvement.
- The pUB110 minus origin functions in multiple bacterial hosts, unlike others described.
Conclusions:
- Plasmid pUB110 employs distinct mechanisms for its plus and minus origins.
- Rolling circle replication is proposed for the plus origin.
- RNA polymerase plays a role in minus origin initiation.
- The broad host range of the pUB110 minus origin offers advantages for genetic engineering.