1a.Objectives (from AD-416)
1. Identify MDV Meq and c-Jun binding sites in the chicken genome.
2. Identify differentially expressed genes following MDV-reactivation in MSB-1 cells.
3. Determine if differentially expressed genes identified in objective 2 also show differentially expression between MD tumors and CD4+ T cells.
4. Identify differentially expressed genes as a function of genetic resistance status or the presence of Meq.
1b.Approach (from AD-416)
1. Utilize chromatin immunoprecipitation (ChIP) using Meq or c-Jun antibodies and determine the DNA sequences via Solexa paired-end reads.
2. Isolate RNA from MSB-1 cells at time 0 and following MDV reactivation, and run on Affymetrix Chicken microarrays.
3. Compare by qRT-PCR the relative expression of key genes between MD tumors and CD4+ T cells.
4. Isolate RNA from line 6 (MD resistant) and line 7 (MD susceptible) CEF at various time before and after infection with either MDV or MDV lacking Meq, and run on Affymetrix Chicken microarrays.
This project is directly linked to Specific Cooperative Agreement 3635-31320-008-26S "Positional Candidate Genes for Resistance to Marek's Disease by Screening for Marek's Disease Virus Meq-Regulated Genes." Marek’s disease, a T cell lymphoma of chickens caused by a pathogenic virus, costs the poultry industry approximately $1 billion worldwide annually in losses and control measures. Thus, understanding how the Marek’s disease virus leads to disease is of both scientific and commercial interest. Meq is the putative cancer-causing viral protein and leads to aberrant expression of chicken proteins. Using molecular biology techniques and high-throughput sequencing, an ARS scientist in East Lansing identified all the targets candidate genes in the chicken genome that Meq might alter. This information can help efforts to develop more effective vaccines or improve genetic resistance, both of which would reduce costs and improve animal welfare. The project is proceeding as planned. This project is monitored by e-mail, telephone calls, and almost weekly face-to-face meeting between the two parties.