Project Number: 1265-32000-086-01
Project Type: Reimbursable

Start Date: Sep 01, 2004
End Date: Aug 31, 2008

Objective:
Develop chicken intestinal microarray chips which contains 10,000 non-redundant EST sequences and investigate the molecular basis for the macrophage-mediated innate immune responses in the gut to Salmonella and Eimeria. Identify the nature of host genes which control the protective immunty against Salmonella and Eimeria by conducting high throughput gene expression assays using chicken intestinal and macrophage chips.

Approach:
Chicken intestinal microarray chips will be first developed by PCR reaction of 10,000 non-redundant intestinal genes selected from chicken intestinal cDNA library. Chickens will be infected with coccidia or salmonella, and RNA samples will be prepared from infected intestinal lymphocytes. Primary macrophages will be prepared from broiler chickens and infected with Eimeria or Salmonella for probe RNA preparation. Molecular basis for the protective innate immune responses in the gut to Salmonella and Eimeria infections will be investigated by carrying out high-throughput gene expression analysis using intestinal and macrophage cDNA microarrays. The role of host immune factors which were identified in the Objective 1 will be validated by real time PCR and northern blot analysis and host factors which are responsible for the intracellular killing of Salmonella and Eimeria will be identified.