焦化废水活性污泥PAH双加氧酶基因多样性分析

Diversity of PAH dioxygenase genes of activated sludge from coking wastewater

为分析焦化废水活性污泥中降解PAH双加氧酶的多样性,利用16Sr DNA-PCR-DGGE方法,以实际焦化废水好氧单元活性污泥总DNA为模板,通过引物对污泥中的双加氧酶基因进行了克隆表达和多样性分析.结果表明,以活性污泥总DNA为模板,利用引物RHD-GN-610F和RHD-GN-916R扩增后有明显的产物,产物大小约为300bp;DGGE指纹图谱显示PCR产物有8条分离条带,丰度分别为2.99%、8.16%、20.75%、28.50%、8.62%、7.26%、10.62%和13.10%;条带经切胶回收PCR扩增后出现明显的扩增产物,TA克隆后成功测试出4条序列,长度分别为305bp,298bp,334bp和294bp,表明焦化废水活性污泥中存在不同降解PAH的RHD酶.这些结果为焦化废水中PAHs的风险评估及其潛在生物降解提供理论基础.

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 by16Sr DNA-PCR-DGGE methodusing total DNA of aerobic activated sludge as the template.The results showed that significant amplification product wasfound using primers RHD-GN-610F and RHD-GN-916R,and the product size was300bp.PCR product was conducted byDGGE analysis and eight separate bands were found,the abundance was2.99%,8.16%,20.75%,28.50%,8.62%,7.26%,10.62%and13.10%,respectively.Obvious amplification products further appeared after strips by gel extraction and PCRamplification,and4sequences were tested successfully by TA cloning,the length of the sequences was305bp,298bp,334bp and294bp,respectively,indicating the presence of different PAH RHD enzymes in coking activated sludge.Theseresults provide a theoretical basis for the risk assessment and potential biodegradation of PAHs.