RESUMO
The gene encoding the beta subunit of the translation initiation factor eIF-2 in the yeast Saccharomyces cerevisiae was mapped by physical methods to the distal part of the left arm of chromosome XVI, adjacent to the HSP90 locus. This assignment was confirmed by genetic linkage data with the GAL4 locus.
Assuntos
Mapeamento Cromossômico , Fator de Iniciação 2 em Eucariotos/genética , Saccharomyces cerevisiae/genética , Genoma FúngicoRESUMO
The gene encoding the beta subunit of the translation initiation factor eIF-2 in the yeast Saccharomyces cerevisiae was mapped by physical methods to the distal part of the left arm of chromosome XVI, adjacent to the HSP90 locus. This assignment was confirmed by genetic linkage data with the GAL4 locus
Assuntos
Mapeamento Cromossômico , Código Genético , Biossíntese de Proteínas , Saccharomyces cerevisiae/genética , Southern Blotting , Genoma FúngicoRESUMO
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.
Assuntos
Proteínas de Bactérias/genética , Glicosilfosfatidilinositóis/genética , Mycobacterium leprae/imunologia , Proteínas de Saccharomyces cerevisiae , Proteínas de Bactérias/imunologia , Proteínas Fúngicas/genética , Genes Fúngicos , Vetores Genéticos , Glicosilfosfatidilinositóis/imunologia , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Mycobacterium leprae/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/imunologiaRESUMO
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
