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| Diversity of PAH dioxygenase genes of activated sludge from coking wastewater |
| MENG Xiao-jun1,2, LI Hai-bo2, SHENG Yu-xing2, CAO Hong-bin2 |
1. School of Tourism and Environment, Ankang University, Ankang 725000, China;
2. Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China |
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Abstract In order to analyze the biodiversity of PAH dioxygenase in activated sludge from a full-scale coking wastewater, primers were used for dioxygenase gene cloning expression and diversity analysis by 16Sr DNA-PCR-DGGE method using total DNA of aerobic activated sludge as the template. The results showed that significant amplification product was found using primers RHD-GN-610F and RHD-GN-916R, and the product size was 300bp. PCR product was conducted by DGGE analysis and eight separate bands were found, the abundance was 2.99%, 8.16%, 20.75%, 28.50%, 8.62%, 7.26%, 10.62% and 13.10%, respectively. Obvious amplification products further appeared after strips by gel extraction and PCR amplification, and 4sequences were tested successfully by TA cloning, the length of the sequences was 305bp, 298bp, 334bp and 294bp, respectively, indicating the presence of different PAH RHD enzymes in coking activated sludge. These results provide a theoretical basis for the risk assessment and potential biodegradation of PAHs.
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Received: 10 May 2016
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