Microbial Genome Sequencing: Sequencing Genomes of the Myoviridae

Tulane University, New Orleans LA

Investigators

Abstract

The project is a collaborative effort between the PI, Jim D. Karam, Ph.D. (Tulane University) and Henry M. Krisch, Ph.D. (CNRS, Toulouse, France). The purpose of the project is to sequence the genomes of up to 8 divergent members of the Myoviridae Family whose virion morphologies resemble phage T4 to varying extents. The genomes of the Myoviridae constitute a rich, and yet largely unexplored, genetic pool for proteins that play a wide variety of biological roles. The wild-type phage strains chosen for analysis infect phylogenetically diverse bacterial hosts that inhabit a variety of ecological niches. They include three coliphages (RB69, RB49 and RB43) that are distantly related to T4, three Aeromonas phages (44RR2.8t, Aeh1, and 25), a Burholderia cepacia phage (phage 42) that also infects some Pseudomonads, and an Acinetobacter phage (phage 133). It is expected that the project will generate a database and experimental tools that will enhance the research activities of evolutionary biologists, geneticists, biochemists, and structural biologists. A cost-effective agreement has been arranged with Integrated Genomics, Inc. (a biotechnology company) to construct genomic libraries for the phage strains of interest and to carry out high-throughput sequencing on these libraries. It is expected that each genome will receive 6X coverage during the high-throughput phase of the project and two subsequent rounds of primer walking to close gaps. Annotated sequences will be made available to the scientific community on the web (GenBank and Integrated Genomics servers) as soon as 6X coverage is obtained and verified. DNA samples and other assistance will be provided to potential collaborators and others in the scientific community. Monthly updates will be placed in the public databases (at GenBank and Integrated Genomics). Also, the project will be used as a backdrop for enhancement of graduate courses at Tulane University via instruction in genomics technology and bioinformatics. This is a Microbial Genome Sequencing Award funded through a collaborative activity between the National Science Foundation and the Department of Agriculture Initiative for Future Agriculture and Food Systems.