水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2014年
3期
454-458
,共5页
DGGE可靠性指数%DNA指纹%变性梯度凝胶电泳%微生物群落结构
DGGE可靠性指數%DNA指紋%變性梯度凝膠電泳%微生物群落結構
DGGE가고성지수%DNA지문%변성제도응효전영%미생물군락결구
DGGE reliable index%DNA fingerprinting%Denaturing gradient gel electrophoresis (DGGE)%Microbial community structure
为了评估DGGE的可靠性，对DGGE条带中回收的DNA片段进行了测序比较分析，并引入了DGGE可靠性指数的概念评价其可靠性。结果显示同一条DGGE条带回收的DNA来自同一属的概率为64.7%，相同位置的DGGE条带可以被认为是同一OTU；不同的DGGE条带回收到类似的DNA序列(16S rDNA V3区差异小于4 bp)的概率为10.5%；DGGE可靠性指数为74.8%。以上结果表明尽管DGGE技术与理论预期存在一定的差距，但是DGGE技术基本能够反映微生物群落的多样性。
為瞭評估DGGE的可靠性，對DGGE條帶中迴收的DNA片段進行瞭測序比較分析，併引入瞭DGGE可靠性指數的概唸評價其可靠性。結果顯示同一條DGGE條帶迴收的DNA來自同一屬的概率為64.7%，相同位置的DGGE條帶可以被認為是同一OTU；不同的DGGE條帶迴收到類似的DNA序列(16S rDNA V3區差異小于4 bp)的概率為10.5%；DGGE可靠性指數為74.8%。以上結果錶明儘管DGGE技術與理論預期存在一定的差距，但是DGGE技術基本能夠反映微生物群落的多樣性。
위료평고DGGE적가고성，대DGGE조대중회수적DNA편단진행료측서비교분석，병인입료DGGE가고성지수적개념평개기가고성。결과현시동일조DGGE조대회수적DNA래자동일속적개솔위64.7%，상동위치적DGGE조대가이피인위시동일OTU；불동적DGGE조대회수도유사적DNA서렬(16S rDNA V3구차이소우4 bp)적개솔위10.5%；DGGE가고성지수위74.8%。이상결과표명진관DGGE기술여이론예기존재일정적차거，단시DGGE기술기본능구반영미생물군락적다양성。
To assess the reliability of DGGE, the V3 region fragments of prokaryotic 16S rDNA sequences were ampli-fied by PCR and separated by DGGE with 40%—70%denaturing gradient in present study, and the differences of DNA sequences retrieved from DGGE bands were compared, and the concept of DGGE reliable index (RI) that was calculated by the equation RI=(Nss-Nsd+Ndd-Nds)/Ns, was introduced. The results showed that the probability of a single band retrieving similar sequences (the sequence diversity of the V3 region of 16S rDNA is less than 4 bp) was 64.7%, and that the bands located at the same level of denaturing gradient could be considered as the same OTU. The probability that different bands retrieve similar sequences was 10.5%. The DGGE reliable index was 74.8%. Therefore, the results of DGGE still reflect the molecular diversity of environmental microbiota, although there was a disparity comparing with theoretical expectations.
