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United States Department of Agriculture

Agricultural Research Service

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Research Project: IDENTIFICATION, CHARACTERIZATION, AND VALIDATION OF GENETIC MUTATIONS INCURRED DURING IN VITRO ATTENUATION OF MAREK’S DISEASE VIRUS

Location: Avian Disease and Oncology Laboratory

Project Number: 3635-32000-016-03
Project Type: Specific Cooperative Agreement

Start Date: Feb 01, 2010
End Date: Dec 15, 2013

Objective:
As described in USDA AFRI-funded grant where Dr. Niikura is a co-PI, we will: 1. Identify and correlate specific genetic and functional changes in the MDV genome that occur during the in vitro attenuation process with virulence. 2. Confirmation of the relevance of the data obtained in objective 1 to the attenuation process.

Approach:
With respect to Objective 1, we will: 1. Serially passed BAC-cloned Marek’s disease virus (MDV) Md5 strain in triplicate and test for virulence. 2. Characterize genetic changes in the MDV genome using next generation sequencing, and correlate the allele frequency results with the level of virulence. 3. Characterize sequence changes that occur during the attention process using next generation sequencing, and correlate the expression differences with the level of virulence. 4. Determine SNP allele frequencies in MD vaccines. With respect to Objective 2, we will: 1. Validate changes identified in objective 1 but introducing defined mutations in our MDV-BAC clone.

Last Modified: 8/1/2014
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