Döring F, Klapper M, Theis S, Daniel H
Institute of Nutritional Sciences, University of Giessen, Germany.
Biochem Biophys Res Commun. 1998 Sep 18;250(2):531-5. doi: 10.1006/bbrc.1998.9342.
The promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (PGAP) was employed to produce the mammalian peptide transporters hPEPT1 and rPEPT2 as models for polytopic transmembrane proteins in the methylotrophic yeast Prichia pastoris. Cells of a recombinant renal peptide transporter (rPEPT2) clone produced constitutively the functional carrier protein. The level of functional expression of rPEPT2 with PGAP varied depending on the carbon source used for cell growth, but was up to five times higher than that obtained with the commonly employed inducible alcohol oxidase 1 promoter (PAOX1). Similar results were obtained for the expression level of the human intestinal peptide transporter hPEPT1 controlled by either PGAP or PAOX1. Therefore, the PGAP seems to be an attractive alternative to PAOX1 for generation of transgenic P. pastoris cells expressing functional mammalian membrane transport proteins at high levels.
利用甘油醛-3-磷酸脱氢酶基因（PGAP）的启动子，在甲基营养型酵母巴斯德毕赤酵母中生产哺乳动物肽转运体hPEPT1和rPEPT2，作为多跨膜蛋白的模型。重组肾肽转运体（rPEPT2）克隆的细胞组成性地产生功能性载体蛋白。使用PGAP时，rPEPT2的功能表达水平因用于细胞生长的碳源而异，但比常用的诱导型醇氧化酶1启动子（PAOX1）获得的表达水平高出五倍。对于由PGAP或PAOX1控制的人肠道肽转运体hPEPT1的表达水平，也获得了类似的结果。因此，对于产生高水平表达功能性哺乳动物膜转运蛋白的转基因巴斯德毕赤酵母细胞而言，PGAP似乎是PAOX1的一个有吸引力的替代方案。
