不同16SrDNA靶序列对DGGE分析活性污泥群落的影响

为探讨不同通用引物扩增16S rDNA靶序列对活性污泥微生物群落分析的影响,更合理的利用变性梯度凝胶电泳(DGGE)技术分析活性污泥样品.从连续流搅拌槽式反应器(CSTR)中获取活性污泥,以3对通用引物341f/534r、968f/1 401r和341f/926r扩增16S rDNA序列,用DGGE分离PCR扩增产物.研究表明采用不同引物对进行DGGE分析时,群落多样性和动态存在显著的差异.341f/534r和968f/1 401r的靶序列分离效果较好,341f/926r的靶序列分离效果较差.引物341f/534r和341f/926r DGGE图谱显示S2和S3相似性高,引物968f/1 401r DGGE图谱显示S1和S2相似性高.由此可见采用不同引物对进行DGGE分析时,群落结构之间的相似性和动态是不一致的.341f/534r的DGGE图谱中条带丰富,多样性最好,968f/1 401r的DGGE图谱次之,341f/926r DGGE图谱条带最少,多样性也较差.因此,在利用DGGE分析活性污泥样品时采用引物341f/534r和968f/1 401r是比较适宜的.

Community of Activated Sludge Based on Different Targeted Sequence of 16S rDNA by Denaturing Gradient Gel Electrophoresis

In order to realize effect of different sets of universal primers on the analysis of microbial community of activated sludge based on targeted sequence of 16S rDNA and employ effectively denaturing gradient gel electrophoresis(DGGE),activated sludge were obtained from the continuous stirred tank reactor(CSTR),16S rDNA fragments were amplified with there primer sets(341f/534r,(968f/1?401r) and 341f/926r),and the diversity and dynamics of microbial communities were investigated by DGGE.The results indicated that community diversity and dynamics was obviously different based different sets of universal primers by DGGE.Separated patterns of the targeted sequence of primer 341f/534r and(968f/1?401r) were better than of(341f/926r.) The similarity of communities between S2 and S3 was high in the DGGE profiles with primer 341f/534r and 341f/926r,the similarity of communities between S1 and S2 was high in the DGGE profiles with primer(968f/1?401r,) it demonstrated that the similarity and dynamics of communities was different each other based different sets of universal primers.In the DGGE profiles, bands and diversity from primer 341f/534r were most,bands and diversity from primer 341f/926r were least.Thereby,while activated sludge was analyzed by DGGE,primers 341f/534r and(968f/1?401r) were more effective than 341f/926r.