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Characterization of a plasmid from moderately halophilic eubacteria
R Fernandez-Castillo1, C Vargas, J J Nieto
1Department of Microbiology, Faculty of Pharmacy, University of Sevilla, Spain.
Journal of General Microbiology
|June 1, 1992
Summary
A novel 11.5 kbp plasmid (pMH1) was isolated from moderate halophilic bacteria, offering antibiotic resistance. Its characteristics make it a promising cloning vector for halophilic organisms.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Moderate halophilic eubacteria inhabit saline environments.
- Plasmids are extrachromosomal DNA elements crucial for bacterial adaptation.
- Characterization of native plasmids is essential for genetic manipulation of extremophiles.
Purpose of the Study:
- To isolate and characterize a novel plasmid from moderate halophilic eubacteria.
- To assess the potential of the isolated plasmid as a cloning vector.
Main Methods:
- Plasmid isolation and transformation into Escherichia coli JM101.
- Restriction mapping and Southern hybridization.
- Determination of antibiotic resistance markers.
Main Results:
- An 11.5 kbp plasmid, designated pMH1, was successfully isolated and transformed.
- Unique restriction sites (EcoRI, EcoRV, ClaI) were identified, facilitating cloning.
- The plasmid confers resistance to kanamycin, tetracycline, and neomycin.
- pMH1 demonstrated stability in E. coli.
Conclusions:
- pMH1 is the first reported plasmid from moderate halophilic eubacteria.
- Its stability, antibiotic resistance, and unique restriction sites make it a valuable tool.
- pMH1 is a strong candidate for developing a cloning vector for moderate halophiles.