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Title: A modified gene trap approach to identify secretory molecules involved in mouse development
Author: Brennan, Jane
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1997
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In an effort to understand the molecular mechanisms that mediate cellular interactions during development I have used a modified gene trap vector, the secretory trap vector (pGT1.8TM), to enrich for insertions in genes encoding secreted and membrane-spanning molecules. The selective property of this vector depends on the presence of a 680 bp fragment containing the transmembrane domain of CD4 fused to the βgeo reporter gene. Secretory trap lines were generated by electroporating pGT1.8TM into CGR8 ES cells. One line generated in this study, JST185 carries an insertion of pGT1.8TM and was selected for detailed characterisation on the basis of its spatially restricted expression pattern during development. The endogenous gene carrying the insertion in JST185 was identified by 5'RACE and cDNAs were isolated. The full length JST185 cDNA contains a single open reading frame which shows homology to a C.elegans gene of unknown function. The hydropathy profiles predict both genes encode the N-terminal signal sequence and ten transmembrane domains. Their intracellular domains contain a tandem array of tetratricopeptide repeats (TPRs), motifs shown to mediate protein-protein interactions. The expression pattern of JST185 during embryogenesis suggests it plays a role in mediating epithelial-mesenchymal interactions during development. Animals homozygous for the mutant allele die shortly after birth due to respiratory failure. The few that survive beyond weaning have breathing difficulties which get progressively worse, these animals die between 3 weeks and 6 months of age. The histology of newborn mutant lungs suggests that they are immature while the lungs of the few surviving homozygotes show gross abnormalities in alveolar architecture and signs of pneumonia which would account for their premature death. In summary JST185 carries a secretory trap insertion in a novel multiple membrane spanning molecule which appears essential for normal lung development.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available