GFP为报告基因的酵母哺乳动物细胞穿梭载体的构建及其在人血管内皮细胞中的表达

为研究酵母作为载体在口服基因治疗及免疫中的作用 ,需要一种能够在酵母中复制而在哺乳动物细胞中表达的穿梭载体 .利用通用载体质粒融合系统 (UPS)构建了一种以GFP为报告基因的新载体 ,以常规的氯化锂法对酿酒酵母进行转化 ,证明该载体能够在酵母中复制 ;以脂质体介导向人血管内皮细胞进行了转染 ,有绿色荧光 ,证明该载体能够在哺乳动物细胞中表达 .所获得的新型的穿梭载体为口服酵母在基因治疗中的应用提供了物质准备

The Construction of Yeast-Mammal Cell Shuttle Plasmid and Expression of Report Gene GFP in HUEVC

A novel shuttle plasmid which can be replicated in yeast and expressed in mammal cell is required in oral gene therapy with yeast as vector to deliver target gene. With GFP as report gene the plasmid was constructed by use of the UPS. By conventional lithium chloride method, the palsmid was transformed into Saccharomyces erevisiae to testify whether it could be replicated in yeast. By transfecting experiments induced by Lipofectamine R, it was testified that the reported gene GFP could be expressed in mammalian cell such as HESVC. The results showed that the novel shuttle plasmid constructed is suitable to be taken as a vector in oral gene therapy.