| 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. |
