PCR-DGGE鉴定短程硝化与反硝化系统的亚硝酸菌

Identification of Nitritebacteria in Short-Cut Nitrifying-and-Denitrifying System by PCR-DGGE technology

现代分子生物学技术PCR-DGGE是一种分析微生物群落的有效工具，可以用于研究生态系统中微生物多样性。首先运用传统方法分离培养亚硝酸菌群，提取DNA后通过PCR-DGGE技术对16S rDNA进行PCR扩增，结合DGGE，鉴定短程硝化与反硝化系统中的亚硝酸菌群。用NCBI-Blast软件将测序结果在Genbank数据库中进行同源性检索，测序结果经检索证实亚硝酸菌优势菌与标准菌株Nitrosomonas eutropha C91的保守性片段有87％的同源性，传统培养方法结合PCR-DGGE测序技术是一种可行且有效的鉴定亚硝酸菌的分析方法。

As a modern molecular technology, PCR-DGGE is a powerful tool for the analysis of microbial communities, which could be used to study the multiformities of microbial communities. In this paper, to study the multiformities of microbial communities of nitritebacteria in Short-Cut Nitrifying-and-Denitrifying System, traditional method was done to isolate and cultivate nitritebactcria communities, then extrude crude DNA , separate and amplify 16S rDNA by PCR and sequence it with DGGE. Homology analysis was made by searching in Genbank with NCBI-Blast. The results show that 87% of nucleotides of the predominance of nitrite bacteria were identical with the conserved fragment of Nitrosomonas eutropha C91. PCR-DGGE technology combined with traditional cultivation method is a feasible and efficient method for identification of nitritebacteria.