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利用位于X染色体上的6-氧嘌呤鸟嘌呤磷酸核糖转移酶(hypoxanthine guenine phosphoribosy ltranstrase HPRT)控制基因缺欠时一般不会将介质中与嘌呤碱基结构相似的6-巯基鸟嘌呤(6-thiogu anine,6-TG)掺入到DNA导致死亡的这一特点,研究了辐射引起的小鼠骨髓造血干细胞HPRT基因突变。由于造血干细胞(CFU-S)在骨髓中的含量约为粒—巨噬系定向干细胞GM-CFC数量的1/10,直接进行CFU-S的HPRT基因突变分析会有相当困难,因此采取了确定GM-CFC受照后是否产生这一突变来回推其祖先CFU-S的HFRT基因突变的存在的方法。实验首先采用RPMI-1640培养液进行正常骨髓GM-CFC的离体软琼脂双层培养,培养体系中含马血清35％,用小鼠肺C·M作CSF,浓度为10％,用6-TG进行选择以确定其最适浓度。实验表明,当骨髓有核细胞数为2×1 0~5/皿时,于37℃含
The use of hypoxanthine guenine phosphoribosyltransferase (HPRT) on the X chromosome to control gene deficiency generally does not involve the 6-thioguanine (6-thioguanine), which is structurally similar to the purine base in the medium , 6-TG) into DNA-induced death, we studied the radiation-induced mutation of HPRT gene in mouse bone marrow hematopoietic stem cells. Since CFU-S is present in the bone marrow at about 1/10 the number of granulocyte-macrophage-derived stem cells GM-CFCs, it is quite difficult to directly analyze the mutation of HPRT gene of CFU-S, GM-CFC after irradiation produced this mutation back and forth to push their ancestors CFU-S HFRT gene mutation exists. In the experiment, first, RPMI-1640 culture medium was used to culture bovine GM-CFC in vitro soft agar. The culture system contained 35% horse serum and the mouse lung C · M was used as CSF in a concentration of 10% TG to choose to determine its optimum concentration. Experiments show that when the number of bone marrow nucleated cells 2 × 10 ~ 5 / dish, at 37 ℃ containing