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Updated: Feb 15, 2026

Genome-wide Purification of Extrachromosomal Circular DNA from Eukaryotic Cells
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Circular permutation engineering improves amplification efficiency of Bst DNA polymerase.

Rong Xiang1, GuangYi Liu2, YanRu Wang1

  • 1School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, China.

International Journal of Biological Macromolecules
|February 13, 2026
PubMed
Summary

Researchers engineered a novel DNA polymerase (CP-G23) for enhanced nucleic acid detection. This polymerase simplifies primer design and eliminates the need for reverse transcriptase in loop-mediated isothermal amplification (LAMP) assays.

Keywords:
Bst DNA polymeraseCircular permutationInhibitor toleranceLoop-mediated isothermal amplification (LAMP)Reverse-transcription LAMP (RT-LAMP)

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostic Assays

Background:

  • Chimeric Bst DNA polymerase (HpStBL) showed promise for nucleic acid detection but faced limitations.
  • Complex primer design and nonspecific amplification hindered broader application of HpStBL in loop-mediated isothermal amplification (LAMP).

Purpose of the Study:

  • To develop improved DNA polymerases for loop-mediated isothermal amplification (LAMP) and reverse-transcription LAMP (RT-LAMP).
  • To create a simplified, rapid nucleic acid detection method with enhanced performance.

Main Methods:

  • Generated four circular permutation (CP) Bst DNA polymerase mutants by altering the Hp47-Sto7d domain of HpStBL.
  • Evaluated DNA synthesis and reverse transcriptase activity of the mutants.
  • Developed a nucleic acid detection system using the CP-G23 mutant and Thermus thermophilus recombinase.

Main Results:

  • Circular permutation altered protein hydrophobicity and DNA affinity, yielding mutants with DNA synthesis and reverse transcriptase activity.
  • Mutant CP-G23 exhibited superior amplification performance, detecting as low as 100 copies/μL Astroviruses plasmid DNA.
  • The CP-G23-based system successfully amplified Astroviruses DNA and SARS-CoV-2 RNA in 45 minutes using a single primer pair, without needing reverse transcriptase.

Conclusions:

  • Circular permutation is an effective strategy for improving DNA polymerase performance in LAMP assays.
  • The novel CP-G23 polymerase and associated detection method offer a simplified, rapid, and sensitive approach for DNA/RNA detection.
  • This method holds significant potential for clinical diagnostics and molecular testing.