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


Autoria(s): Han Yifan; Mo Zhaolan; Mao Yunxiang; Xiao Peng; Li Jie; Hao Bin; Guo Dongsheng
Data(s)

01/08/2009

Resumo

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 by 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 by 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.

Identificador

http://ir.qdio.ac.cn/handle/337002/2915

http://www.irgrid.ac.cn/handle/1471x/166112

Idioma(s)

英语

Fonte

Han Yifan; Mo Zhaolan; Mao Yunxiang; Xiao Peng; Li Jie; Hao Bin; Guo Dongsheng.Construction and characterization of a fosmid library for pathogenic bacterium Vibrio anguillarum,CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,2009,27(3):519-526

Palavras-Chave #Limnology; Oceanography #end-sequencing #fosmid library #PCR screening #Vibrio anguillarum
Tipo

期刊论文