嗜水气单胞菌气溶素基因的克隆与序列分析.PDFVIP

微生物学通报 MAY 20, 2008, 35(5): 700~704 Microbiology © 2008 by Institute of Microbiology, CAS tongbao@ 研究报告 嗜水气单胞菌气溶素基因的克隆与序列分析 * 李寿崧 郭立新 江树勋 陈文炳 邵碧英 ( 350001) 摘 要: 以嗜水气单胞菌BZ 和NK 分离株的 DNA 为模板, 采用 PCR 技术, 扩增气溶素基因(aerA) 的 DNA 片段, 将其克隆到 pMD18-T 载体上。通过序列测定, 分析结果表明：所克隆的 1393 bp 片段为 aerA 部分序列, 编码产生 464 个氨基酸。BZ 与 NK 之间 aerA 核苷酸同源性为 97.6%, 氨 基酸同源性为 98.3%, 与其它分离物核苷酸同源性为 71.6%~97.5%, 氨基酸同源性为 68.0%~98.9%。利用邻接法构建了 aerA 分子树状图, 树状图分析表明：气单胞菌属各分离物聚为 三支, 其中嗜水气单胞菌各菌株之间关系密切, 被聚类为同一支。 关键词: 嗜水气单胞菌, 气溶素基因, 克隆, 序列分析 Cloning and Nucleotide Sequence Analysis of Aerolysin Gene of Aeromonas hydrophila * LI Shou-Song GUO Li-Xin JIANG Shu-Xun CHEN Wen-Bing SHAO Bi-Ying (Fujian Entry-exit Inspection and Quarantine Bureau of P.R.C, Fuzhou 350001) Abstract: The aerolysin genes (aerA) of BZ and NK isolates were cloned and sequenced. The sequence analysis showed that the partial aerA of BZ and NK isolates consisted of 1393 bp, encoding a protein of 464 amino acids. The similarity of nucleotide and amino acid sequences of aerA between BZ and NK isolates was 97.6% and 98.3% respectively. The nucleotide sequence of aerA of BZ strain exhibited 71.6% to 97.5% homology with other Aeromonas isolates, and the amino acid sequence exhibited 68.0% to 98.9% homology. The phylogenetic tree based on aerA nucleotide sequences from Aeromonas isolates was constructed with neighbor-joining method. It showed that there were three branches of aerolysin genes, and a close rel