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Published on: May 2, 2018
Curing Both Virulent Mega-Plasmids from Bacillus anthracis Wild-Type Strain A16 Simultaneously Using Plasmid
Dongshu Wang1, Zhiqi Gao1, Huagui Wang1
1State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
Abstract:
Plasmid-cured derivative strains of Bacillus anthracis are frequently used in laboratory studies. Plasmid incompatibility, which does not increase the risk of chromosomal mutation, is a useful method for plasmid curing. However, in bacteria containing multiple plasmids, it often requires the sequential introduction of multiple, specific incompatibility plasmids. This lengthy process renders the traditional plasmid incompatibility method inefficient and mutation-prone. In this study, we successfully cured plasmids pXO1 and pXO2 from B. anthracis A16 simultaneously using only one recombinant incompatible plasmid, pKORT, to obtain a plasmid-free strain, designated A16DD. This method may also be useful for the simultaneous, one-step curing of multiple plasmids from other bacteria, including Bacillus thuringiensis and Yersinia pestis.
Insights
Researchers developed a faster method to remove plasmids from Bacillus anthracis using a single incompatible plasmid. This streamlined process efficiently creates plasmid-free bacterial strains for laboratory use.
Area of Science:
- Microbiology
- Bacterial Genetics
- Molecular Biology
Background:
- Plasmid-cured Bacillus anthracis strains are essential for laboratory research.
- Traditional plasmid curing via incompatibility is inefficient for multiple plasmids.
- Sequential introduction of incompatibility plasmids is time-consuming and mutation-prone.
Purpose of the Study:
- To develop an efficient, one-step method for simultaneous plasmid curing in Bacillus anthracis.
- To obtain a plasmid-free Bacillus anthracis A16 strain using a novel approach.
Main Methods:
- Utilized a single recombinant incompatible plasmid, pKORT.
- Simultaneously cured both pXO1 and pXO2 plasmids from B. anthracis A16.
- Obtained a plasmid-free derivative strain, designated A16DD.
Main Results:
- Successfully achieved simultaneous, one-step curing of pXO1 and pXO2 from B. anthracis A16.
- Generated a plasmid-free Bacillus anthracis strain (A16DD).
- Demonstrated the efficiency of the pKORT plasmid for plasmid elimination.
Conclusions:
- The developed method offers a significant improvement over traditional plasmid curing techniques.
- This one-step, simultaneous plasmid curing approach is efficient and reduces mutation risk.
- The method holds potential for simultaneous multi-plasmid curing in other bacterial species like Bacillus thuringiensis and Yersinia pestis.
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