Construction of suppression subtractive cDNA library and analysis of differentially expressed genes in the spleen of the chicken

To screen the differentially expressed genes related to anti-disease activity in chicken,a suppression subtractive hybridization(SSH) was employed to construct the SSH cDNA library in the spleen by using Dagu chicken and commercial Hyland brown chicken at 20 weeks of age.The inserted fragment sizes at 600 bp and 760 clones were verified by PCR,of which 663 clones were used to perform dot blotting hybridization and 531 clones were obtained.Subsequently,100 clones were randomly picked and sequenced.37 differentially expressed genes were isolated by homologous BLAST search.Besides 5 unknown function genes,the other 32 genes implicated in general disease resistance-related and immunity,specific disease resistance-related,cell signaling molecules,membrane protein and transcriptional factor genes,and so on.Four differentially expressed genes probably impacting anti-disease activity of chicken were identified using semi-quantitative RT-PCR method.The Construction of SSH cDNA Library was expected to lay the foundation for isolating and cloning the novel genes from chicken spleen,which potentially involved in spleen performance,and for further investigating the functions and the regulatory mechanisms of these genes during anti-disease activity.