Title Monitoring 4-Chlorobiphenyl-Degrading Bacteria in Soil Microcosms by Competitive Quantitative PCR
Author Soo Youn Lee, Min Sup Song, Kyung Man You, Bae Hoon Kim, Seong Ho Bang 1 , In Soo Lee 2 , Chi Kyung Kim 3 , and Yong Keun Park*
Address Graduate School of Biotechnology, Korea University, Seoul 136-701, Korea; 1 Department of Biology, Hanseo University, Seosan 356-706, Korea; 2 Department of Microbiology, Hannam University, Daejeon 300-791, Korea; 3 Department of Microbiology, Chungbuk National University, Cheongju 360-763, Korea
Bibliography Journal of Microbiology, 40(4),274-281, 2002,
Key Words 4-chlorobiphenyl degrading bacteria, soil microcosms, competitive quantitative PCR
Abstract The competitive quantitative PCR method targeting pcbC gene was developed for monitoring 4-chlorobiphenyl(4CB)-degrading bacteria, Pseudomonas sp. strain DJ-12, in soil microcosms. The method involves extraction of DNA from soil contaminated with 4CB, PCR amplification of a pcbC gene fragment from the introduced strain with a set of strain-specific primers, and quantification of the electrophoresed PCR product by densitometry. To test the adequacy of the method, Pseudomonas sp. Strain DJ-12 was introduced into both contaminated and non-contaminated soil microcosms amended with 4CB. Pseudomonas sp. strain DJ-12 was monitored and quantified by a competitive quantitative PCR in comparison with 4CB degradation and the result was compared to those obtained by using the conventional cultivation method. We successfully detected and monitored 4CB-degrading bacteria in each microcosm and found a significant linear relationship between the number of 4CB-degrading bacteria and the capacity for 4CB biodegradation. The results of DNA spiking and cell-spreading experiments suggest that this competitive quantitative PCR method targeting the pcbC gene for monitoring 4Cbdegrading bacteria appears to be rapid, sensitive and more suitable than the microbiological approach in estimating the capacity of 4CB biodegradation in environmental samples.
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