Expression of Mycobacterium leprae 18-kDa antigen in yeast in a GPI-anchored form.
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.
Amorim
A G
AG
Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, São Paulo, Brasil.
Cardoso-de-Almeida
M L
ML
Carrington
M
M
Morga
D P
DP
Riezman
H
H
Zerbini
L F
LF
Piestun
V S
VS
Castilho-Valavicius
B A
BA
D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D005822:Genetic Vectors; D017261:Glycosylphosphatidylinositols / Q000235:genetics* / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D009166:Mycobacterium leprae / Q000235:genetics / Q000276:immunology*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000276:immunology; D029701:Saccharomyces cerevisiae Proteins
C067864:18 kDa protein, Mycobacterium leprae; D001426:Bacterial Proteins; D005656:Fungal Proteins; C066709:GAS1 protein, S cerevisiae; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D029701:Saccharomyces cerevisiae Proteins