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Transformation of Acholeplasma laidlawii with streptococcal plasmids pVA868 and pVA920
1Department of Comparative Medicine, University of Alabama, Birmingham 35294.
Abstract:
One limitation with studying mycoplasma genetics is the lack of cloning vectors. Studies were undertaken to determine whether streptococcal plasmids could replicate in Acholeplasma laidlawii, for the purpose of identifying potential vectors. Plasmids pVA868 and pVA920 contain the same origin of replication and tetracycline resistance determinant. pVA920 also contains an erythromycin resistance determinant not present in pVA868. A. laidlawii was transformed with plasmids pVA868 (13.7 kb) and pVA920 (12.2 kb), and isolated from the transformants were deletion derivatives of the parent plasmids having sizes of 3.7 and 10.3 kb, respectively. The tetracycline and erythromycin resistance markers functioned in A. laidlawii, and the deletion derivatives may be useful for development of mycoplasma vectors. However, difficulties may arise due to plasmid instability.
Insights
Researchers explored using streptococcal plasmids to create cloning vectors for Acholeplasma laidlawii genetics. They found that resistance markers functioned, and modified plasmids show potential for mycoplasma vector development despite instability.
Area of Science:
- Molecular Biology
- Microbiology
Background:
- Studying mycoplasma genetics is hindered by the absence of suitable cloning vectors.
- Streptococcal plasmids were investigated as potential candidates for replication in Acholeplasma laidlawii.
Purpose of the Study:
- To assess the replicability of streptococcal plasmids in Acholeplasma laidlawii.
- To identify potential cloning vectors for mycoplasma genetic studies.
Main Methods:
- Transformation of Acholeplasma laidlawii with streptococcal plasmids pVA868 and pVA920.
- Isolation and characterization of deletion derivatives from transformed plasmids.
- Verification of antibiotic resistance marker function in Acholeplasma laidlawii.
Main Results:
- Deletion derivatives of pVA868 (3.7 kb) and pVA920 (10.3 kb) were successfully isolated from transformed Acholeplasma laidlawii.
- Tetracycline and erythromycin resistance markers were functional in the host.
- Identified deletion derivatives show promise as mycoplasma vectors.
Conclusions:
- Streptococcal plasmids can replicate in Acholeplasma laidlawii, yielding functional resistance markers.
- The identified deletion derivatives represent potential tools for developing mycoplasma cloning vectors.
- Plasmid instability may present challenges in the practical application of these vectors.