Ultra-High Throughput Sequencing for Discovery and Characterization of Viral Genomes in Fire Ants
2010 Annual Report
1a.Objectives (from AD-416)
Use a metagenomics approach to identify new viruses infecting the fire ant Solenopsis invicta and subsequently use genomic tools and resources to study the biology (distribution, pathogenicity, etiology, molecular evolutionary history) of several of these viruses.
1b.Approach (from AD-416)
1) Perform a large-scale EST study using 454 pyrosequencing technology to identify potential viruses infecting fire ants using bioinformatics tools and manual annotation of genes.
2) Confirm that newly discovered viruses occur within S. invicta and are not contaminants.
3) Sequence the entire genomes of newly discovered viruses.
4) Use an existing cDNA microarray to determine the host response to viral infections by comparing and identifying genes differentially expressed between virus-infected and uninfected fire ants.
5) Sequence multiple genomes of a single virus and multiple alleles of host genes that respond to viral infection and examine their molecular evolution.
This project is related to sub-objective 1.1 of this in-house project: Perform large-scale EST study to significantly increase the breadth and depth of transcriptome coverage using 454 pyrosequencing technology.
Four cDNA libraries were constructed and were then sequenced using 454 pyrosequencing technology. Formal analyses of resulting transcriptome data for viruses using bioinformatic tools is underway.
Monitoring: The ARS scientist monitored Progress on this project by means of frequent email communications, telephone communications, and visits to the collaborator’s laboratory.