从pHIG53质粒内切下人白细胞介素-2(IL-2)cDNA基因, 并经中间质粒pSP72转换成与pDOR-neo载体相匹配的酶切位点, 然后将IL-2cDNA定向连接入pDOR-neo载体, 构建成功人IL-2逆转录病毒载体, 经脂质体导入人骨肉瘤细胞系Ma中, 经G418筛选后测转基因肿瘤细胞培养上清中IL-2表达量, 每1×105细胞24hIL-2表达量为50~800U, 为骨肉瘤的基因治疗创造条件.
The human IL-2 cDNA was cleaved from pHIG53 and inserted in the intermediate vector pSP-72 which has the restriction sites comptible with clone site of the expression vector pDOR-neo, then IL-2 cDNA was directionally inserted into the retroviral vector pDOR-neo to construct a eukaryotic expression vector. The vector was trans feeted into human osteosarcoma cell line by lipofectin. The IL-2 was bioassayed in the culture of tumor cell after selection of G418. The production of IL-2 was 50~800 U per 1×105 cells every 24h, which makes it possible to carry out osteosarcoma gene therapy.
郑强,范清宇,殷剑宁,惠宏襄.人白细胞介素-2基因在人骨肉瘤细胞系内表达[J].生物化学与生物物理进展,1996,23(5):467-470
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