摘要
用重组人β－珠蛋白基因及其增强子的逆转录病毒载体转染φ－２细胞及ＰＡ３１７细胞，分离出产病毒粒子的φ－２和ＰＡ３１７克隆，用这两种细胞克隆进行乒乓上清感染，得到能产生高滴度逆转录病毒载体的ＰＡ３１７细胞克隆，其滴度达５．９×１０＾６ＣＦＵ／ｍｌ。
Transferring β-globin gene and its enhancer into human hematopoietic cells is the basisfor applying β-thalassemia gene therapy in clinical practice.We isolated ecotropic virus pro-ducing clones and amphotropic virus producing clones by using a replication-defective retro-virus vector containing β-globin gene and its enhancer to transfect ecotropic packaging cellline -2 and amphotropic packaging cell line PA317.Then by ping-ponging supernatant in-fection between ecotropic producer clone and amphotropic producer clone, we obtained hightiter amphotropic PA317 producer clones with the highest titer up to 5.9×106 CFU/ml.Hu-man mononuclear bone marrow cells were infected repeatly with this high titer virus vectorunder stimulation of hematopoietic growth factors IL-3,IL-6 and SCF,Southern blot hy-bridization analysis showed that β-globin gene and its enhancer had been integrated into thegenome of human hematopoietic cells.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1996年第5期353-357,共5页
Acta Academiae Medicinae Sinicae
基金
"863"基金
关键词
β
珠蛋白
基因
逆转录病毒
载体
骨髓单核细胞
globin gene
retrovirus vector
human mononuclear bone marrow cells
gene transfer
gene therapy
