应用PCR-DGGE技术分析福寿螺胃肠道菌群

The Flora of the Pomacea canaliculata Gastric and Intestinal Contents with PCR-DGGE

为了探究福寿螺消化纤维素的生理机制,本研究采用PCR-DGGE技术分析了福寿螺胃肠内容物的菌群结构。无菌条件下采集福寿螺胃、肠内容物,提取细菌总DNA;以通用引物扩增细菌16S r DNA V3区;DGGE电泳,回收、克隆、测定DGGE优势条带。结果表明,雌、雄福寿螺胃肠菌群结构相同,均为22种细菌;10个明显条带的序列分别与气单胞菌属(Aeromonas sp.)、希瓦氏菌属(Shewanella sp.)、粘质沙雷菌(Serratia marcescens)、肠杆菌属(Enterobacter)、铁单胞菌属(Ferrimonas sp.)和5个不可培养的细菌的16S r DNA V3区序列高度相似。分离到2株细菌,其中1株为腐败希瓦氏菌。

In order to explore the digestive physiological mechanism of fiber, the florae of the P.canaliculata gastric and intestinal contents were determined with PCR- DGGE. The analysis process were as follows：collecting the gastric and intestinal contents in sterile conditions, extracting the bacterial total genomic DNA,amplifying the 16 S r DNA V3 with universal primers, DGGE electrophoresis; recycling, cloning and sequencingthe selected bands. The results showed that the flora of the male and female P. canaliculata gastric andintestinal contents were the same, containing 22 bacteria; the sequences of the ten distinct bands were highlysimilar with Aeromonas sp, Shewanella sp., Serratia marcescens, Enterobacter, Ferrimonas sp. and otheruncultured bacteria. Two kinds of bacteria were separated, one of them was Shewanella putrefaciens.