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16S与gyrB基因联合建树快速鉴定海水中的一株地衣芽胞杆菌
傅奇, 林俊杰, 甘美裕, 卢源钦, 肖玉娟
厦门华厦学院
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,目前常用的方法为增加测序基因数,并分别建树,但容易出现树间结构冲突、测序基因多的问题。课题组从海水中筛选得到一株杆菌,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,分别将其16S和gyrB基因序列合并,采用Paup* 4.0构建进化树。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:  16S  gyrB  进化树  地衣芽胞杆菌
DOI:
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 with phylogenetic tree based on combination of 16S and gyrB geneXIAO Yu-Juan, LIN Junjie, GAN Meiyu, LU Yuanqin, FU Qi *
fuqi, linjunjie, ganmeiyu, luyuanqin, xiaoyujuan
Xiamen Huaxia University
Abstract:
In the identification of bacteria with highly similar conserved sequences, 16S rDNA/RNA sequence only for blast or construction of phylogenetic tree is usually not accurate enough. Currently, the commonly used method is to sequencing more genes and establish more trees separately, which bring about more work and structural conflicts about phylogenetic trees. A strain of bacillus from seawater was isolated, 16S and gyrB genes of the strain were amplified with universal primers and sequenced. By blast in genebank, strains with both similar 16S and gyrB genes were selected. The 16S and gyrB genes sequences of these strains were combined separately and a phylogenetic tree based on combined sequences was constructed using Paup* 4.0. The strains belonging to the same specie in the tree are well polymerized in one branch, and the self-expanding values of the branches are higher than 98, and the classification structure is accurate. The isolated strain is polymerized with Bacillus licheniformis in the tree, and the self-expanding value is 100. It was verified by physiological and biochemical tests that the strain belong to specie of Bacillus licheniformis. The identification result obtained by construction of phylogenetic tree based on combination of 16S and gyrB gene was accurate, rapid and simple.
Key words:  16S  gyrB  phylogenetic tree  Bacillus licheniformis
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