Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A Novel Nucleic Lateral Flow Assay for Screening phaR-Containing Bacillus spp.

  • Wint, Nay Yee (Department of Biochemistry, Faculty of Pharmacy, Mahidol University) ;
  • Han, Khine Kyi (Department of Pharmacology, Faculty of Pharmacy, Mahidol University) ;
  • Yamprayoonswat, Wariya (Chulabhorn Graduate Institute, Chulabhorn Royal Academy) ;
  • Ruangsuj, Pattarawan (Chulabhorn Graduate Institute, Chulabhorn Royal Academy) ;
  • Mangmool, Supachoke (Department of Pharmacology, Faculty of Pharmacy, Mahidol University) ;
  • Promptmas, Chamras (Department of Biomedical Engineering, Faculty of Engineering, Mahidol University) ;
  • Yasawong, Montri (Department of Biochemistry, Faculty of Pharmacy, Mahidol University)
  • Received : 2019.07.21
  • Accepted : 2019.10.24
  • Published : 2021.01.28
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Abstract

Polyhydroxyalkanoate (PHA) synthase is a key enzyme for PHA production in microorganisms. The class IV PHA synthase is composed of two subunits: PhaC and PhaR. The PhaR subunit, which encodes the phaR gene, is only present in class IV PHA synthases. Therefore, the phaR gene is used as a biomarker for bacteria that contain a class IV PHA synthase, such as some Bacillus spp. The phaR gene was developed to screen phaR-containing Bacillus spp. The phaR screening method involved two steps: phaR gene amplification by PCR and phaR amplicon detection using a DNA lateral flow assay. The screening method has a high specificity for phaR-containing Bacillus spp. The lowest amount of genomic DNA of B. thuringiensis ATCC 10792 that the phaR screening method could detect was 10 pg. This novel screening method improves the specificity and sensitivity of phaR gene screening and reduces the time and cost of the screening process, which could enhance the opportunity to discover good candidate PHA producers. Nevertheless, the screening method can certainly be used as a tool to screen phaR-containing Bacillus spp. from environmental samples.

Keywords

References

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