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Construction of a shuttle vector for use between Pasteurella multocida and Escherichia coli
M M Bills1, J M Medd, R J Chappel
1Department of Microbiology, Monash University, Clayton, Victoria, Australia.
Plasmid
|November 1, 1993
Summary
Researchers created a new shuttle vector, pBAC64, for genetic engineering in both Pasteurella multocida and Escherichia coli. This tool facilitates cloning and manipulation across these bacterial species.
Area of Science:
- Microbiology
- Molecular Biology
- Bacterial Genetics
Background:
- Pasteurella multocida is a significant bacterial pathogen.
- Genetic manipulation of P. multocida can be challenging.
- A shuttle vector allows for replication and manipulation in two different host organisms.
Purpose of the Study:
- To construct a novel shuttle vector for use in both Pasteurella multocida and Escherichia coli.
- To facilitate genetic studies and manipulation in P. multocida.
Main Methods:
- Isolation of a small plasmid from P. multocida.
- Construction of the shuttle vector pBAC64 by incorporating P. multocida and E. coli elements.
- Characterization of the vector's replication and selection capabilities.
Main Results:
- The shuttle vector pBAC64 was successfully constructed.
- pBAC64 contains a functional origin of replication from P. multocida.
- The vector includes an antibiotic resistance gene effective in P. multocida and features of the E. coli vector pUC18, including a multiple cloning site and lacZ' gene for screening.
Conclusions:
- The pBAC64 shuttle vector enables efficient cloning and manipulation in both P. multocida and E. coli.
- This vector is a valuable tool for molecular biology research involving these two bacterial species.