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本文利用逆转录病毒介导的基因转移方法，将人多药耐药基因ｍｄｒ－１导入小鼠造血细胞，并对转基因造血细胞的耐药性及移植特性进行了初步研究。含人ｍｄｒ－１ｃＤＮＡ的重组逆转录病毒载体质粒，经脂质体介导的基因转移方法导入包装细胞ＰＡ３１７，再经秋水仙素筛选。获得产逆转录病毒的包装细胞，其病毒滴度可达１．２×１０５ｃｆｕ／ｍｌ。在含小鼠骨髓贴壁细胞层的长期培养体系中，利用逆转录病毒上清对造血细胞进行转染。通过抗性ＣＦＵ－ＣＭ检测，其基因转移效率可达７．０％。将转基因造血细胞移植经照射预处理的小鼠，可重建受体小鼠造血功能。小鼠造血重建后４个月，其骨髓造血细胞与外周血细胞可检测到人ｍｄｒ－１ｃＤＮＡ，且骨髓造血细胞对秋水仙素仍保留抗性
In this paper, mdr-1, a multidrug resistance gene, was introduced into mouse hematopoietic cells by retroviral-mediated gene transfer. The drug resistance and transplantion characteristics of transgenic hematopoietic cells were also studied. Recombinant retroviral vector containing human mdr-1 cDNA was introduced into packaging cell line PA317 by liposome-mediated gene transfer method and then selected by colchicine. To obtain retroviral packaging cells, the virus titer up to 1.2 × 105cfu / ml. In a long-term culture system containing a mouse bone marrow adherent cell layer, hematopoietic cells are transfected with a retrovirus supernatant. Through the detection of resistant CFU-CM, the gene transfer efficiency can reach 7.0%. The hematopoietic function of recipient mice can be reconstructed by transfection of transgenic hematopoietic cells transplanted with irradiated pre-treated mice. Four months after hematopoietic reconstitution in mice, human mdr-1 cDNA can be detected from bone marrow hematopoietic cells and peripheral blood cells, and myeloid hematopoietic cells remain resistant to colchicine