Construction and characterization of a fosmid library for pathogenic bacterium Vibrio anguillarum  

作　　者：韩一凡 莫照兰 茅云翔 肖鹏 李杰 郝斌 郭东升 

机构地区：[1]Ocean University of China [2]Institute of Oceanology,Chinese Academy of Sciences

出　　处：《Chinese Journal of Oceanology and Limnology》2009年第3期519-526,共8页中国海洋湖沼学报（英文版）

基　　金：Supported by the National Basic Research Program of China (973 Program)(No 2006CB101803);the High Technology Research and development Program of China (863 Program)(No 2006AA6100310);the National Natural Science Foundation of China (No 30871935)

摘　　要：Vibrio anguillarum is a common bacterial pathogen in fish.However,little is known about its pathogenic mechanism,in part,because the entire genome has not been completely sequenced.We constructed a fosmid library for V.anguillarum containing 960 clones with an average insert size of 37.7 kb and 8.6-fold genome coverage.We characterized the library by end-sequencing 50 randomly selected clones.This generated 93 sequences with a total length of 57 485 bp covering 1.4% of the whole genome.Of these sequences,58(62.4%) were homologous to known genes,30(32.3%) were genes with hypothetical functions,and the remaining 5(5.3%) were unknown genes.We demonstrated the utility of this library by PCR screening of 10 genes.This resulted in an average of 6.2 fosmid clones per screening.This fosmid library offers a new tool for gene screening and cloning of V.anguillarum,and for comparative genomic studies among Vibrio species.Vibrio anguillarum is a common bacterial pathogen in fish. However, little is known about its pathogenic mechanism, in part, because the entire genome has not been completely sequenced. We constructed a fosmid library for V. anguillarum containing 960 clones with an average insert size of 37.7 kb and 8.6-fold genome coverage. We characterized the library by end-sequencing 50 randomly selected clones. This generated 93 sequences with a total length of 57 485 bp covering 1.4% of the whole genome. Of these sequences, 58 （62.4%） were homologous to known genes, 30 （32.3%） were genes with hypothetical functions, and the remaining 5 （5.3%） were unknown genes. We demonstrated the utility of this library by PCR screening of 10 genes. This resulted in an average of 6.2 fosmid clones per screening. This fosmid library offers a new tool for gene screening and cloning of V. anguillarum, and for comparative genomic studies among Vibrio species.

关 键 词：end-sequencing fosmid library PCR screening Vibrio anguillarum 

分 类 号：S941[农业科学—水产养殖]