利用PCR-DGGE技术分析肉牛阴道菌群结构

发布时间：2018-04-02 16:12

本文选题：肉牛　切入点：黄体期和卵泡期　出处：《中国兽医学报》2016年01期

【摘要】：利用PCR-DGGE技术明确肉牛阴道菌群结构并比较黄体期和卵泡期肉牛阴道菌群结构差异。选择7头肉牛,分别采集黄体期和卵泡期阴道分泌物,提取细菌总DNA,采用巢式PCR分别对细菌16SrDNA的V3区进行扩增,将PCR产物进行变性梯度凝胶电泳(denatured gradient gel electrophoresis,DGGE)。对DNA指纹图谱特异性条带切胶回收并进行克隆测序,通过BLAST与已知序列对比,鉴定细菌菌种,分析比较黄体期和卵泡期肉牛阴道菌群结构。结果显示:健康肉牛阴道内存在阴道气球菌(Aerococcus vaginalis str.)、绿色气球菌(Aerococcusviridans str.)、睡眠嗜血杆菌(Haemophilussomnus str.)、多动物链球菌(Streptococcus pluranimalium str.)、嗜冷杆菌(Psychrobactermarincolastr.)、大肠杆菌(Escherichia coli O157:H7str.)和鞘氨醇单胞菌(Sphingomonasroseiflavastr.)等。
[Abstract]:The structure of vaginal flora of beef cattle was determined by PCR-DGGE technique and the differences between luteal phase and follicular phase were compared.Seven beef cattle were selected to collect vaginal secretions from luteal phase and follicular stage, and total bacterial DNA was extracted. The V3 region of bacterial 16SrDNA was amplified by nested PCR, and the PCR product was denatured gradient gel electrophoresism DGGEN by denaturing gradient gel electrophoresis.The specific band cut gum of DNA fingerprint was recovered and cloned and sequenced. The bacterial species were identified by comparing BLAST with known sequences and the structure of vaginal flora of beef cattle in luteal phase and follicular stage was analyzed and compared.The results showed that in the vagina of healthy beef cattle, there were Aerococcus vaginalis str., Aerococcus viridans str., Haemophilus somnus str., Streptococcus pluranimalium str., Psychrobacter sp., Escherichia coli O157: H7str.)And Sphingomonas sphingomonas rosei flavastr.Etc.
【作者单位】：吉林农业大学动物科技学院;
【基金】：吉林省现代肉牛产业技术体系项目(201534) 吉林省重大科技攻关专项(20140203016NY) 吉林省发改委产业技术研发专项(2013779) 国家自然科学基金资助项目(31172227)
【分类号】：S852.61

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