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Development of a catabolically significant genetic probe for polycyclic aromatic hydrocarbon-degrading Mycobacteria in soil

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Abstract

A gene probe for the detection of polycyclic aromatic hydrocarbon (PAH) induced nidB and nidA dioxygenase genes has been designed from Mycobacteria JLS, KMS, and MCS. The probe detects a catabolic gene involved in the initial steps of PAH biodegradation in mycobacteria. The gene probe is comprised of three PCR primer sets designed to detect the genes that code for two subunits of the PAH induced dioxygenase enzyme within PAH-degrading mycobacteria. The probe was built by combining three primer sets with a DNA extraction procedure that was designed to lyse the gram-positive mycobacteria cells while in the soil matrix and remove PCR inhibitors. The probe was tested on PAH contaminated soils undergoing bioremediation through landfarming and uncontaminated soils from the same site. The PAH gene probe results demonstrate that the dioxygenase genes can be detected in soils. Sequencing the nidA and nidBPCR products verified that the genes were detected in soil. Comparisons of the sequences obtained from the soil probe to seven known nid gene sequences from various PAH-degrading mycobacteria showed between 97 and 99% nucleotide matches with the nidB gene and 95 and 99% matches with the nidA gene.

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Authors and Affiliations

  1. North Wind, Inc., 1425 Higham Street, Idaho Falls, ID, 83402, USA

    Kevin Hall

  2. Department of Biology, Utah State University, 5305 Old Main Hill, Logan, UT, 84322-8200, USA

    Charles D. Miller & Anne J. Anderson

  3. Department of Biological and Irrigation Engineering, College of Engineering, Utah State University, 5305, Old Main Hill, Logan, UT, 84322-8200, USA

    Ronald C. Sims

  4. Utah Water Research Laboratory, College of Engineering, Utah State University, 5305 Old Main Hill, Logan, UT, 84322-8200, USA

    Darwin L. Sorensen & Ronald C. Sims

Authors
  1. Kevin Hall
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  2. Charles D. Miller
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  3. Darwin L. Sorensen
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  4. Anne J. Anderson
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  5. Ronald C. Sims
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Correspondence to Kevin Hall.

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Hall, K., Miller, C.D., Sorensen, D.L. et al. Development of a catabolically significant genetic probe for polycyclic aromatic hydrocarbon-degrading Mycobacteria in soil. Biodegradation 16, 475–484 (2005). https://doi.org/10.1007/s10532-004-5669-y

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  • DOI: https://doi.org/10.1007/s10532-004-5669-y

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