16SrDNA克隆文库法分析拜城油鸡盲肠细菌多样性研究

发布时间：2018-06-28 05:01

本文选题：拜城油鸡 + 盲肠　；参考：《家畜生态学报》2017年02期

【摘要】：为了研究拜城油鸡盲肠细菌物种多样性,本研究采用基于16SrDNA基因序列系统发育分析的免培养方法进行细菌物种多样性分析,以盲肠内容物样品总DNA为模板,利用细菌通用引物扩增16SrDNA基因,构建16SrDNA基因克隆文库,并对文库中的插入序列进行RFLP分析。结果表明:随机挑选的252个克隆为阳性克隆,阳性率为65.7%,经过RFLP筛选出95个不同图谱的重组克隆并测序,比对分析后发现:拜城油鸡盲肠细菌分属于厚壁菌门(Firmicutes)(65.26%)、拟杆菌门(Bacteroidetes)27.37%)和变形菌门(Proteobacteria)(4.21%)。在纲水平有梭菌纲(Clostridia)、拟杆菌纲(Bacteroidia)和ε-变形菌纲(Epsilonproteobacteria)相对含量最多,分别占总菌的64.21%、26.32%和4.21。在科水平上以瘤胃菌科(Ruminococcaceae)(21.05%)、毛螺菌科(Lachnospiraceae)(20%)和理研菌科(Rikenellaceae)(12.63%)相对含量最多,为优势科。在属水平上仅有31.58%的序列可以鉴定到属,Lachnoclostridium的相对含量最多,占总菌的12.63%,其次为另枝菌属(Alistipes)和瘤胃梭菌属(Ruminiclostridium)。拜城油鸡盲肠内容物中的细菌组成具有丰富的多样性,并有新细菌分类单位的潜在资源,值得进一步进行开发研究。
[Abstract]:In order to study the species diversity of cecum bacteria in Baicheng fowl, the bacterial species diversity was analyzed by using 16s rDNA gene phylogenetic analysis method. The total DNA of cecum contents was used as template. 16s rDNA gene was amplified by bacterial universal primers and the 16s rDNA gene clone library was constructed. The inserted sequence of 16s rDNA gene was analyzed by RFLP. The results showed that 252 clones randomly selected were positive clones with a positive rate of 65.7. 95 recombinant clones with different profiles were screened by RFLP and sequenced. The results showed that the cecum bacteria belonged to Firmicutes (65.26%), Bacteroidetes (27.37%) and Proteobacteria (4.21%). The relative contents of Clostridia, Bacteroidia and Epsilon proteobacteria were the most, accounting for 64.21% and 4.21% of the total bacteria, respectively. The relative contents of Ruminococcaceae (21.05%), Lachnospiraceae (20%) and Rikenellaceae (12.63%) were the dominant family. At the generic level, only 31.58% of the sequences could be identified as the relative content of Lachnoclostridium, accounting for 12.63% of the total bacteria, followed by (Alistipes) and Ruminiclostridium. The bacterial composition in the cecum of Baicheng fowl is rich in diversity and potential resources of new bacteria taxonomic units, so it is worthy of further development and research.
【作者单位】：塔里木大学动物科学学院新疆生产建设兵团塔里木畜牧科技重点实验室;
【基金】：兵团塔里木畜牧科技重点实验室开放课题(HS201205) 国家自然科学基金(31160521) 塔里木大学校长基学校长基金(TDZKGG201504)
【分类号】：S831;S852.6

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