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Updated: Jun 19, 2026
14:26
Genome-wide Purification of Extrachromosomal Circular DNA from Eukaryotic Cells
Published on: April 4, 2016
Cloning of circular DNAs from microorganisms using a novel plasmid capture system
Jong Yul Roh1, Yong Wang, Qin Liu
1Department of Agricultural Biotechnology, Seoul National University, Republic of Korea.
Molecular Biotechnology
|October 20, 2009
Summary
The improved Plasmid Capture System (PCS-LZ) enables cloning of large DNA molecules (30-150 kb). This powerful tool aids genomic analysis and mutagenesis in microorganisms, including invertebrate pathogens.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Plasmid capture systems (PCS) are essential for cloning and manipulating circular double-stranded DNA.
- Efficient cloning of large DNA fragments remains a challenge in molecular biology.
Purpose of the Study:
- To develop an improved PCS (PCS-LZ) for cloning large DNA molecules (30-150 kb).
- To demonstrate the utility of PCS-LZ in genomic analysis and mutagenesis of various DNA sources.
Main Methods:
- The PCS-LZ donor, containing a mini-F replicon and kanamycin resistance marker flanked by Tn7 ends, was used.
- In vitro transposition transferred the donor elements into target plasmid DNAs.
- Replication in E. coli and blue/white screening (lacZ expression) were employed for selection.
Main Results:
- PCS-LZ successfully cloned circular DNAs from diverse sources, including Bacillus thuringiensis plasmids, Cotesia glomerata bracovirus genome segments, and Autographa californica nucleopolyhedrovirus genomes.
- The system demonstrated efficiency in handling large DNA molecules within the 30-150 kb range.
Conclusions:
- PCS-LZ is a powerful and versatile tool for genomic analysis and targeted mutagenesis.
- The system is particularly valuable for applications involving microorganisms, including invertebrate pathogens.
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