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From IGGI

About IGGI

The International Glossina Genome Initiative (IGGI) was established through WHO/Tropical Disease Research and Training (TDR) in 2004 with the ultimate goal to impact on tsetse control by generating a fully annotated whole genome sequence for Glossina – the vector for Human African Trypanosomiasis.

Mission statement

The overall mission for IGGI is to enhance trypanosomiasis control by developing genomic resources which will:

• Promote the understanding of Glossina biology including host-pathogen interactions, genetics of vector competence and population genetics to support vector control efforts
• Stimulate the development of new vector control initiatives including potential targeted insecticide development, host seeking studies leading to improved trap and target design, indirect transgenic approaches via symbionts
• Lead to an increase in the size and impact of the Glossina research community
• Help generate a research resource development and access facility
• Promote training and capacity building in disease endemic countries

Members

• Dr Serap Aksoy, Yale, USA (Acting Chair)
• Dr Mike Lehane, Liverpool, UK
• Dr Neil Hall, TIGR, USA
• Dr Matt Berriman, Wellcome Trust Sanger Institute, UK
• Dr Patrick Wincker,Genoscope, Paris
• Dr Masahira Hattori, RIKEN, Japan
• Dr Winston Hide, SANBI, South Africa
• Dr Elliot Krafusr, Iowa, USA
• Dr Atway Msangi, TTRI, Tanzania
• Dr Loyce Okedi, LIRI, Uganda
• Dr Philippe Solano, IRD-CIRDES, Burkina Faso
• Dr Jan van den Abbeele, ITMA, Belgium
• Dr Todd Taylor, RIKEN, Japan
• Dr Lucien Manga, VBC, Division of Health Environments & Sustainable Development, WHO AFRO, Congo
• Dr Deborah Kioy, TDR, WHO
• Dr Ayoade Oduola, TDR, WHO
• Dr Yeya Touré, TDR, WHO

Projects

Glossina morsitans morsitans genome project

Whole genome shotgun sequencing

Whole genome shotgun sequencing to 3-fold genome coverage will take place at the WTSI. Small insert sequencing libraries have been prepared from a single fly to polymorphism in the genome assembly. Currently (19th Oct. 07), the project is at ~1-fold coverage.

BAC library construction and sequencing

The Tsetse fly (Glossina morsitans morsitans) BAC library was constructed by Garnet Navarro and Andrew Stuart in Chris Amemiya's laboratory at the Benaroya Research Institute at Virginia Mason. This library was designated VMRC-29. Tsetse fly specimens were obtained from Dr. Serap Aksoy, Yale University School of Public Health. These specimens were reared on antibiotic-containing media in order to reduce gut microflora. Nuclei were prepared from whole pupae (~50) using a dounce homogenizer and several rounds of centrifugation. The nuclei were then embedded in Incert agarose and high molecular weight DNA was prepared in situ in the agarose. Generation of the library closely followed the cloning approach developed in Pieter de Jong's laboratory . Partial-digest EcoRI fragments from the appropriate size fraction were cloned into the pCCBACE1 vector (Epicentre Technologies). The ligation products were then transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library has been arrayed into 144 384-well microtiter dishes and has subsequently been gridded onto 22x22cm nylon high-density colony filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones (equivalent to 48 "384-well" dishes), stamped in duplicate. Analysis of 125 randomly selected BACs via pulsed field gel electrophoresis of NotI-digested DNA indicated that the median insert size was around 152 kb and the average insert size was 138 kb (the descrepancy is due to a small fraction of smaller insert-containing clones in the library, i.e., clones < 100 kb).

The library filters and the BAC clones can be purchased. Information at http://bacpac.chori.org/library.php?id=354

Three BACs have been finished at WTSI and their sequences can be downloaded from ftp://ftp.sanger.ac.uk/pub/pathogens/Glossina/morsitans/

Five BACs were finished at RIKEN. Nearly 20,000 BAC clones were end-sequenced at the University of Tokyo in collaboration with RIKEN and Hokkaido University.

Fosmid library construction and sequencing

A Fosmid library is under construction at WTSI using the same fly as used for the small-insert libraries. Fosmids have a tightly constrained insert size of 40 kb, thus end sequencing of cloned inserts produces paired sequencing reads with a fixed 40 kb distance between them that greatly facillitates assembling the shotgun sequencing data into large supercontigs or scaffolds.

Nearly 10,000 fosmid clones were end-sequenced at RIKEN.

EST library construction and sequencing

EST library sequencing - WTSI

• Midgut cDNA library (Lehane lab) normalised by the Soares laboratory, Iowa. The midgut project has been published.
• Salivary gland cDNA library (Aksoy lab), normalised by the Soares laboratory, Iowa.
• Head cDNA library (Lehane lab). 2700 ESTs have been sequenced and are available
• Reproductive organs cDNA library (Aksoy lab). 3438 ESTs have been sequenced and are available

EST sequencing - Yale & TIGR

• Fat body cDNA library (Aksoy lab), normalised by the Soares laboratory, Iowa. The fat body project has been published.

Other EST sequencing

• Head and Antennae libraries (G. pallipedes, G. palpalis, G. tachnoides) have been produced (Masiga and Berriman labs) and are in testing.

cDNA sequencing

5'-end sequencing and 3'-end sequencing of 49,398 full length cDNA clones from various tissues including whole body male and female adults, pupae and larva, were completed by Dr. Junichi Watanabe (University of Tokyo) in collaboration with RIKEN.

Glossina palpalis palpalis sequencing project

Documents

Glossina White Paper 2007

Get involved

Please contact: Serap Aksoy: serap.aksoy_at_yale.edu