Use of suppressive subtractive hybridization to identify differentially expressed genes in ayu (Plecoglossus altivelis) associated with Listonella anguillarum infection

Chang H Li; Jiong Chen; Yu H Shi; Xin J Lu

doi:10.1016/j.fsi.2011.06.011

Use of suppressive subtractive hybridization to identify differentially expressed genes in ayu (Plecoglossus altivelis) associated with Listonella anguillarum infection

Fish Shellfish Immunol. 2011 Sep;31(3):500-6. doi: 10.1016/j.fsi.2011.06.011. Epub 2011 Jun 22.

Authors

Chang H Li¹, Jiong Chen, Yu H Shi, Xin J Lu

Affiliation

¹ Faculty of Life Science and Biotechnology, Ningbo University, Ningbo City, People's Republic of China.

PMID: 21712095
DOI: 10.1016/j.fsi.2011.06.011

Abstract

Suppressive subtractive hybridization (SSH) was employed to identify differentially expressed genes in ayu (Plecoglossus altivelis) associated with Listonella anguillarum infection. 800 random clones were selected from forward and reverse subtractive libraries and 787 were successfully sequenced. After assembling, 105 contigs and 414 singletons were finally obtained, some of which were immune-related genes. A real-time quantitative PCR (RT-qPCR) analysis of the expression patterns of 28 transcripts showed that the false-positive rate was approximately 7.1%. Furthermore, Wap65-2 was overexpressed in Escherichia coli, purified and used for antiserum preparation. Western blot analysis revealed that serum Wap65-2 of ayu significantly increased after bacterial infection, suggesting that it was a positive acute-phase protein (APP).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Fish Diseases / metabolism
Fish Diseases / microbiology*
Gene Expression Profiling / veterinary*
Gene Expression Regulation / immunology
Gram-Negative Bacterial Infections / metabolism
Gram-Negative Bacterial Infections / veterinary*
Listonella*
Osmeriformes*