傅奇, 林俊杰, 甘美裕, 卢源钦, 肖玉娟
厦门华厦学院环境与公共健康学院, 福建 厦门361024

摘要:
在保守序列高度相似的细菌鉴定中，单独使用16S rDNA/RNA序列进行比对和构建进化树通常无法准确鉴定到种，需要增加测序基因数并对多基因进行分析。为实现快速鉴定，课题组对16S与gyrB基因联合建树的方法进行了研究，将海洋来源的一株杆菌，分别用通用引物扩增16S和gyrB基因并测序，在GeneBank进行序列比对后，选择各菌种保藏中心16S和gyrB基因均相似的菌株，取16S和gyrB基因序列，采用Paup*4.0构建进化树。使用16S与gyrB拼接序列构建的进化树中属于同一种的菌株均很好的聚合在一枝，种间分枝自展值均高于98，分类结构准确，筛选得到的杆菌与地衣芽胞杆菌（Bacillus licheniformis）聚合在一枝，自展值为100，鉴定为地衣芽胞杆菌。经生理生化试验验证，该菌株与地衣芽胞杆菌特征完全一致，使用16S和gyrB基因联合建树得到的鉴定结果准确且快速简便。
关键词:16SgyrB进化树地衣芽胞杆菌(Bacillus licheniformis)
DOI：10.11759/hykx20191018002
分类号:Q939
基金项目:福建省中青年教师教育科研项目（科技类）（JAT170821）；福建省大学生创新创业训练计划项目（201812709001）；福建省本科高校教育教学改革研究项目（FBJG20180124）

Identification of a Bacillus licheniformis strain from seawater using a phylogenetic tree based on 16S and gyrB gene sequences
FU Qi, LIN Jun-jie, GAN Mei-yu, LU Yuan-qin, XIAO Yu-juan
College of Environment and Public Health, Xiamen Huaxia University, Xiamen 361024, China
Abstract:
For identifying bacteria with highly similar conserved sequences, 16S rDNA does not provide adequate resolution for comparative sequence analysis; therefore, more DNA sequences and analysis models are required for classification. To develop a rapid identification method, phylogenetic trees based on 16S and gyrB gene sequences were investigated. A strain of bacillus was isolated from seawater, and the 16S and gyrB genes of the strain were amplified with universal primers and sequenced. Using BLAST in GenBank, other strains with similar 16S and gyrB genes were selected. Phylogenetic trees based on 16S and gyrB genes were constructed using PAUP* 4.0. In the tree constructed using sequences of both 16S and gyrB genes, strains belonging to the same species were clustered together, the self-expanding values of the branches were higher than 98, and the classification structure was accurate. The isolated strain polymerized with Bacillus licheniformis in the phylogenetic tree, and the self-expanding value was 100. Physiological and biochemical tests verified that the strain belongs to the species B. licheniformis. Thus, the identification of this strain of B. licheniformis obtained by construction of a phylogenetic tree based on the combination of 16S and gyrB genes was accurate, rapid, and simple.
Key words:16SgyrBphylogenetic treeBacillus licheniformis





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