Introduction - The TIGR Brugia malayi Genome Project

B. malayi is, with Wuchereria bancrofti, a causative agent of lymphatic filariasis. These parasites are endemic in over 100 countries and place 20% of the world’s population at risk of infection. Disease syndromes include acute lymphangitis, pulmonary eosinophilia and chronic disfiguring elephantiasis.

The goal of this project is to determine 98% of the genomic sequence of B. malayi (TRS strain), analyze and annotate the data and provide access to the sequence information and analysis. Sequences generated from this project will be deposited into the GSS division of GenBank according to our data release policy. We will not attempt closure of the complete sequence of the B. malayi genome. This project is being carried out in collaboration with Dr. Alan Scott, of Johns Hopkins University, Dr. Jeremy Foster and Dr. Barton Slatko, both of New England Biolabs, and in close coordination with the Filarial Genome Network

Brugia malayi microfilaria
	Brugia malayi L3 larvae coming out of a mosquito proboscis.

Sequencing methodology

The approach proposed here is of a whole genome shotgun (WGS) strategy that should enable us to obtain 98% of the DNA sequence of B. malayi in two years.

The specific steps are:

1. Prepare two sheared DNA libraries from adult worms (male and female) B. malayi gDNA. The first library, with a 2kb average insert size, will provide the bulk of the sequence coverage. End sequences from the second library, at about 15-20 kb insert size, will be valuable for ordering of the contig groups and independent verification of overall genome structure.
2. Sequence both ends of approximately 8,000 BAC clones from a BAC library that is currently under construction in collaboration with Dr. Peter de Jong at Children's Hospital Oakland Research Institute. The BAC ends paired markers will be subsequently used to construct a scaffold during genome assembly and provide independent verification of sequence assembly.
3. Assemble the genomic sequences of B. malayi from the collection of random sequence fragments.