摘要
将人促红细胞生成素（ｈＥＰＯ）、人粒细胞集落刺激因子（ｈＧ－ＣＳＦ）、细胞程序死亡抑制基因（ＢＣＬ－２）重组逆转录病毒质粒ＤＮＡ转染病毒包装细胞ＰＡ３１７后，经Ｇ４１８抗性筛选，其单克隆细胞株培养上清液均具有感染ＮＩＨ３Ｔ３及大鼠原代成纤维细胞、成肌细胞并形成Ｇ４１８抗性克隆的能力。其感染细胞的染色体ＤＮＡＰＣＲ鉴定结果均为阳性，表明其染色体中均成功地整合了目的基因；感染的靶细胞均能表达外源基因编码蛋白，表明已成功地建立了逆转录病毒介导的基因转移技术。
Recombinant hEPO,hG-CSF and hBCL-2retroviral plasmids were introduced intovirus-pnckaging cell line PA317 by means of calcium phosphate DNA coprecipitation. G418resistant clones were obtained after 2 weeks G418 screening,which could produce retrovirus intheir coi1ditioned inedium. The retrovirus could transfect NIH3T3 cell line,primary rat fibroblastsand mvoblasts.Genomie DNA PCR identification with specific hEPO,hG-CSF primers showedtliat the tdrget geneS had integrated into the chromosomes of those transfected target cells;CFU-E bioassay resuits indicated that the transfected gene(hEPO cDNA) had expressed in targetcells.All the results above showed that retroviral-mediated gene transfer technology had been es-tablished in our laboratory.
出处
《军事医学科学院院刊》
CSCD
北大核心
1995年第3期222-226,共5页
Bulletin of the Academy of Military Medical Sciences
关键词
逆转录病毒
基因转移
红细胞生成素
retrovirus
gene transfer
erythropoietin
granulocyte colony-stimulating factor
