中文摘要：

建立可稳定表达结核分枝杆菌HSP65-hIL-2融合蛋白的稳定转染P815细胞系。在阳离子脂质体作用下,将HSP65-hIL-2真核表达质粒转染与BALB/c遗传背景一致的P815细胞（H-2^d）。G418筛选阳性克隆,RT-PCR和间接免疫荧光法检测目的蛋白的转录和表达。阳性克隆细胞经RT-PCR检测到HSP65-hIL-2融合基因特异性的mRNA表达;用鼠抗人的IL-2mAb进行间接免疫荧光检测,可在转染的P815细胞浆中观察到较强的绿色特异性荧光,而未转染细胞则为阴性。成功获得稳定表达HSP65-hIL-2融合蛋白的稳定转染细胞系,为其疫苗的CTL研究提供了合适的靶细胞。

英文摘要：

To establish the stably transfected P815 cell line expressing the fusion protein HSP65-hIL-2, the eukaryotic expression plasmid pcDNA-HSP65-hIL-2 was stably transfected into P815 cells （H-2d） using the cationic liposome, whose genetic background was identical to that of BALB/c mice. The positive transfected cells were selected by G418, the specific mRNA of HSP65-hIL-2 was detected by RT-PCR and the expression of the fusion protein in the transfected cells was detected by indirect immunofluorescence technique. The result showed that the specific mRNA of HSP65-hIL-2 could be detected and strong specific green immunofluorescence existed in the cytoplasm of the transfected P815 cells. The stably transfected P815 cell line expressing the fusion protein HSP65-hIL- 2 is constructed suceessfully, which provided target cells for the CTL experiment for the vaccine evaluation related with HSP65-hIL-2 fusion protein.