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Expression of Mycobacterium leprae 18-kDa antigen in yeast in a GPI-anchored form
A G Amorim1, M L Cardoso-de-Almeida, M Carrington
1Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, São Paulo, Brasil.
Abstract:
The 18-kDa protein from Mycobacterium leprae is a major target for the immune response in leprosy. We have developed a system to express this antigen in yeast as a fusion protein with the C-terminal region of the yeast membrane protein GAS1, which would render the recombinant protein anchored to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. Cells lacking the GAS1 gene and transformed with the hybrid 18-kDa-GAS1 construct express a polypeptide that reacts with an 18-kDa-specific monoclonal antibody. In addition, these cells react with an alpha-CRD antibody after GPI-PLC treatment. The non-transformed cells are negative. These data indicate that our system may be suitable for the expression of foreign proteins in yeast in a GPI-anchored form.