Amorim A G, Cardoso-de-Almeida M L, Carrington M, Morga D P, Riezman H, Zerbini L F, Piestun V S, Castilho-Valavicius B A
Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, São Paulo, Brasil.
Braz J Med Biol Res. 1994 Mar;27(3):623-6.
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.
麻风分枝杆菌的18 kDa蛋白是麻风病免疫反应的主要靶点。我们开发了一种系统，可在酵母中表达该抗原，使其作为与酵母膜蛋白GAS1的C端区域融合的蛋白，这将使重组蛋白通过糖基磷脂酰肌醇（GPI）锚定在质膜上。缺乏GAS1基因并转化了杂交18-kDa-GAS1构建体的细胞表达一种与18-kDa特异性单克隆抗体发生反应的多肽。此外，这些细胞在经GPI-PLC处理后与α-CRD抗体发生反应。未转化的细胞呈阴性。这些数据表明我们的系统可能适用于在酵母中以GPI锚定形式表达外源蛋白。
