International Mammalian Genome Society


The 13th International Mouse Genome Conference
October 31-November 3, 1999

Table of Contents * Structure * Bioinformatics * Sequence * Mapping * New Tools * Gene Discovery * Developmental * Mutagenesis * Functional Genomics

I10 Mapping of Transcripts from the Developing Pancreas

Oxford Group: Lorraine Eley, Alison Hugill, Lorraine Southam, Eiifon Herbert, Roger D.Cox: Medical Research Council, Harwell, Oxfordshire, OX11 ORD

EU Consortium:

Paris Group: Philip Avner1,2, Isabelle Poras1, Gabor Gyapy1, Cecile Fizames,1 Nunchanard Chianniluchai,1 William Saurin1, Jean Weissenbach1. 1Genoscope-Centre National de Sequencage, 2 rue Gaston Cremieux, 91006 Evry, France; 2Unite de Genetique Moleculaire Murine, Institut Pasteur, 25 rue du Dr Roux, 75015 Paris, France.

Milano Group: Giacomo Manenti: Instituto Nazionale Tumori, Via G. Venezian I, 20133 Milano, Italy.

Berlin Group: Patricia Ruiz, Michael Wiles, Hans Lehrach: Max-Plank Institute for Molecular Genetics, 14195 Berlin, Germany.

London Group: Rosa Beddington, Sally Dunwoodie, Laboratory of Mammalian Development, National Institute for Medical Research, London, Mill Hill, UK

Cambridge Group: Patricia Rodriguez: European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD, UK

Diabetes mellitus results from the loss of islets of Langerhans b-cells. In type II diabetes islet cells proliferate in response to insulin resistance and it is the loss of this compensatory mechanism or its ability to meet demand that leads to overt diabetes. Transcriptional profiling and mapping of islet expressed genes during islet formation will aid in our understanding of islet biology and may provide candidate genes for diabetes. At embryonic day 16.5 clusters of endocrine cells are present and at embryonic day 17.5 these clusters associate to form islets of Langerhans. Pancreatic islets from 17.5 and 18.5 day embryonic pancreas were isolated using a Ficoll gradient. Using the same method the clusters were isolated from 16.5 day embryos. A directional cDNA library in lTriplEX2 vector was constructed from mouse islet total RNA using a SMARTTM cDNA library construction kit (CLONTECH Palo Alto, CA, USA). The clones were picked using a Biopick into 384 well microtitre plates and then gridded onto nylon membranes using a gridding robot (Genomics solutions). The libraries were characterised by probing the nylon membranes with islet specific genes and exocrine specific genes to determine contamination from this fraction. For each library 18,432 cDNA clones were picked and are currently being sequenced. The number of clones currently sequenced from the 18.5, 17.5 and 16.5 d libraries are 240, 96, 192 respectively. Preliminary sequence data indicates that the libraries are very complex with low redundancy. In the 18.5d library 12.5% of the sequences are not related to any other sequence in the database. In order to map sequences from these islet libraries they are first searched against the EBI mouse cluster database and single sequences from each cluster selected for mapping. Sequences are then searched against the EBI allocation database and ESTs that have not been previously mapped are placed on the mouse radiation hybrid panel T31 using the EU consortiums framework map. Library characterisation, sequencing results and initial mapping data for these libraries will be presented.

 


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