| 秦翠丽，杨若岚，李松彪，李云飞，牛明福，宫强.基于16S rDNA序列4株鸡源芽孢杆菌的分子鉴定[J].广东农业科学,2014,41(22):130-134 |
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基于16S rDNA序列4株鸡源芽孢杆菌的分子鉴定 |
| Molecular identification of four strains of Bacillus from broiler based on 16S rDNA sequences |
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| DOI： |
| 中文关键词: 芽孢杆菌 16S rDNA 分子鉴定 |
| 英文关键词: Bacillus 16S rDNA molecular identification |
| 基金项目:新乡市泰弘生物有限公司委托项目（2477）；河南科技大学SRTP项目（2013119） |
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| 中文摘要: |
| 对分离自肉鸡肠胃的4 株芽孢杆菌R1、R2、S1、H1的16SrDNA序列进行PCR 扩增并测定其核酸序列，在NCBI中通过Nucleotide BLAST 查找其同源序列，应用DNAstar的MegAlign软件中的Jotun Hein、Clustal V、Clustal W3种方法进行序列差异和同源性分析，分别使用Mega5.0软件中的最大似然法、邻接法、最小进化法、最大简约法构建系统发育树。序列差异和同源性分析结果显示：由Jotun Hein 法可知，H1和R2与Bacillus subtilis DSM10 同源性最高，达到100%：R1与B. vallismortis DSM11031 同源性最高，达到99.7%。；H1与B. subtilis DSM10 的同源性为99.9%。由Clustal V法可知，H1与R2同源性最高，达到99.6%；R1与S1同源性最高、为98.7%。由Clustal W法可知，H1与R2同源性最高、为99.7%，R1与S1为99.4%。采用4 种方法构建的系统发育树基本一致，可以确立4 株鸡源芽孢杆菌的分类地位，R1、S1与B. subtilis BPRIST009、B. subtilis LXB3、B. vallismortis DSM11031 之间关系最为亲近；R2、H1与B.subtilis CICC10076、B. subtilis DSM10 之间关系最为亲近。 |
| 英文摘要: |
| 16S rDNA sequences of 4 Bacillus R1, R2, S1, H1, which were isolated from broiler intestines, were amplified by PCR method. Homologous sequences were searched for by Nucleotide BLAST in NCBI. Three methods (Jotun Hein, Clustal V, Clustal W) from MegAlign software of DNAstar respectively were used to analyze the homology differences of sequence. Phylogenetic trees were constructed by using the methods of Maximum Likelihood (ML), Neighbor-Joining (NJ),Minimum Evolution (ME) and Maximum Parsimony (MP), respectively from Mega5.0. The results showed that the homologies of H1 and R2 were both 100% with B. subtilis DSM10 by Jotun Hein method. And the sequence of R1 showed 99.7% homology with B. vallismortis DSM11031. It was also found that the homology between H1 and B. subtilis DSM10 was up to 99.9%. Through the Clustal V method, homology analysis indicated that homology between H1 and R2 was 99.6%, and the homology between R1 and S1 was up to 98.7%. Additionally, the results showed that the homology between H1 and R2 was 99.7% and the homology between R1 and S1 was up to 99.4% by Clustal W method. The phylogenetic trees were basically consistent based on the four methods. The taxonomic status of these four Chicken Bacillus could be confirmed. As R1 and S1 were most similar with B. subtilis BPRIST009, B. subtilis LXB3 and B. vallismortis DSM11031, R2 and H1 were most similar with B. subtilis CICC10076 and B. subtilis DSM10. |
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