致病性鳗弧菌金属蛋白酶基因克隆及序列分析  

Cloning and Sequencing of Metalloprotease Gene of A Pathogenic Vibrio Anguillarum

作　　者：陈吉祥[1] 李筠[1] 王祥红[1] 杜宗军[1] 于德华[1] 纪伟尚[1] 徐怀恕[1] 

机构地区：[1]青岛海洋大学海洋生命学院,青岛266003

出　　处：《高技术通讯》2002年第6期106-110,共5页Chinese High Technology Letters

基　　金：国家自然科学基金 ( 39870 581);中国博士后科学基金资助项目

摘　　要：从我国山东沿海发病的鲈鱼 (Lateolabraxjaponicus)分离到一株致病性鳗弧菌(Vibrioanguillarum)W 1 ,该菌的胞外蛋白酶活性为 42 2 6u/ml,部分纯化的胞外蛋白酶对海水养殖大菱鲆鱼有一定的毒性。应用PCR扩增 ,从鳗弧菌W 1染色体DNA扩增出一条长约 1 .92 5kb的特异性PCR产物 ,DNA序列分析表明 :克隆的片段含有完整的金属蛋白酶基因阅读框 ,编码 61 1个氨基酸残基的蛋白质 ,该金属蛋白酶基因与一株致病性鳗弧菌蛋白酶基因的核苷酸及氨基酸序列同源性为 1 0 0 % ,而与解蛋白弧菌 (V .proteolyticus)、创伤弧菌(V .vulnificus)、霍乱弧菌(V .cholerae)、斑点气单胞菌 (Aeromonaspunctata)、嗜水气单胞菌(Aeromonashydrophila)的氨基酸序列同源性分别为 73 %、70 %、69%、5 3 %、5 1 %。A pathogenic Vibrio anguillarum strain W 1 was originally isolated from diseased seabass ( Lateolabrax japonicus ) in China. Its extra cellular protease activity is 4226u/ml. The partially purified metalloprotease is toxic to turbot. The metalloprotease gene was cloned by PCR amplification from its chromosomal DNA. The nucleotide sequence was determined. The ORF of the gene consists of 1836 bp, which encodes a polypeptide of 611 amino acids. The nucleotide sequence is highly identical to that of a Vibrio anguillarum strain NB10. The similarities of the amino acid sequence of the metalloprotease with those of V. proteolyticus, V. vulnificus, V. cholerae, Aeromonas punctata, Aeromonas hydrophila are 73%, 70%, 69%, 53%, 51% respectively.

关 键 词：基因克隆 鳗弧菌 金属蛋白酶 毒性 基因序列 细菌病 鱼 

分 类 号：Q785[生物学—分子生物学]