应用扩增性rDNA限制性酶切片段多态性分析技术研究竹鼠盲肠细菌菌群的多样性  

Diversity of Cecal Bacterial Flora in Bamboo Rat (Rhizomyidae) Analyzed by Amplified Ribosomal DNA Restriction Analysis Technology

作　　者：周建波[1] 姜亚[1] 宁俊平[1] 曹晓燕[1] 黄怡[1] 王士长[1] 

机构地区：[1]广西大学动物科学技术学院,南宁530005

出　　处：《动物营养学报》2013年第10期2504-2508,共5页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：中柬越英国际合作项目(DelPHE ROUND 5;DelPHE project 852)

摘　　要：本试验应用扩增性rDNA限制性酶切片段多态性分析(ARDRA)技术研究竹鼠盲肠细菌菌群的多样性。利用免培养的分子生物学技术,从竹鼠盲肠内容物中提取细菌的总DNA,用细菌通用引物F27/R1492扩增出细菌16S rDNA的基因,构建16S rDNA基因文库。采用HaeⅢ和HhaⅠ2种限制性内切酶对阳性克隆子的扩增产物进行酶切,挑选不同的操作分类单元(OTU)测序,通过Blast在线比对,分析竹鼠盲肠细菌菌群的多样性。结果表明:竹鼠盲肠中的细菌菌群大部分是未培养或未知的菌株,在分类学上具有潜在意义的被选菌株,而且存在可能和脂肪代谢有关的微生物群体。同时也有乳酸菌属、芽孢杆菌属、梭形杆菌属和多黏类芽孢杆菌属的分布。从结果中看出,芽孢杆菌属、梭形杆菌属和2个未知菌株的克隆数都大于20个,是优势菌株。This experiment was conducted to study the diversity of cecal bacterial flora in bamboo rat （Rhizomyidae） analyzed by amplified ribosomal DNA restriction analysis （ARDRA） technology.The culture-free technology of molecular biology was used to extract microbial total DNA from the cecum of bamboo rat.The bacteria universal primers F27/R1492 were used to amplify bacteria genes of 16S rDNA so as to build up gene library of 16S rDNA.Two kinds of restriction enzymes,HaeⅢ and Hha Ⅰ,were used to digest amplification products of positive clones,and then different operational taxonomic units （OTU） sequencing were picked up.Through Blast comparing online,the diversity of cecal bacterial flora in bamboo rat was analyzed.The results showed that the most microbes in cecum of bamboo rat were uncultured or unknown strains which were the selected strains of potential significance in taxonomy,and there were microorganism groups related to lipid metabolism.Meanwhile,the distribution of Lactobacillus,Bacillus,Fusiformis and Paenibacillus polymyxa also existed.In conclusion,Bacillus,Fusiformis and two unknown strains whose clone numbers are more than 20 are dominant strains.

关 键 词：竹鼠 16S RDNA ARDRA 克隆 

分 类 号：S811.3[农业科学—畜牧学]