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Cloning and sequencing of the cdna coding for two golgi autoantigens ; functional analysis by site-directed mutagenesis

Objective



The first objective of the work will be the cDNA cloning of the genes encoding two autoantigens specific for the Golgi complex, p210 and p230, by immunological screening of two cDNA expression libraries.
The sequence of these clones will be analysed in order to characterize the structure, to understand the function of the proteins and to know wether they are included in a new family of proteins associated to the Golgi apparatus that are preferent targets in certain autoimmune diseases. Fragments of the proteins will be subcloned into a bacterial expression vector in order to overproduce the peptides and to obtain antibodies against p210 and p230. Finally, clones encoding wild type or mutants p210 or p230 will be transfected into eucaryotic cells in order to study the domains involved in the localization and interactions of these proteins in the cell.

Call for proposal

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Coordinator

UNIVERSIDAD DE SEVILLA
EU contribution
No data
Address
S/N,Profesor Garcia Gonzalez S/N
41012 SEVILLA
Spain

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Total cost
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Participants (1)