REGIONAL ASSIGNMENT OF THE GENE FOR DIPHTHERIA-TOXIN SENSITIVITY USING SUBCHROMOSOMAL FRAGMENTS IN MICROCELL HYBRIDS
REGIONAL ASSIGNMENT OF THE GENE FOR DIPHTHERIA-TOXIN SENSITIVITY USING SUBCHROMOSOMAL FRAGMENTS IN MICROCELL HYBRIDS
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DOI：
10.1007/bf00285879
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发表时间：
1987-01-01
期刊：
影响因子：
1.6
通讯作者：
OKADA, Y
中科院分区：
文献类型：
--
作者：
HAYES, H;KANEDA, Y;OKADA, Y
Human. times.在小鼠肝癌细胞和人之间构建了抗G418的小鼠微细胞杂交体。小鼠全细胞杂交体，其仅含有完整的人5号和22号染色体，并具有整合的neor基因。其中，微细胞杂种BG 15产生了四个亚克隆，BG 15 -4、BG 15 -6、BG 15 -7和BG 15 -9，它们含有不同大小的人5号染色体互补序列。BG 15 -6含有完整的人5号染色体，BG 15 -7含有缺失的人5号染色体（5 pter-q22），PG 15 -4和BG 15 -9含有人5号染色体部分之间的易位（pter-qter？和pter-q23）和小鼠染色体。Southern DNA印迹分析表明，人二氢叶酸还原酶（DHFR）基因存在于所有四个亚克隆，而人同源的v-fms基因存在于BG 15 -4和15-6，但不存在于BG 15 -7和15-9。BG 15 -4、15-6和15-9对白喉毒素敏感，只有BG 15 -7对白喉毒素有抗性。我们使用这些微细胞杂交进一步限制白喉毒素敏感性基因在人类5号染色体q23区域的区域定位。
Human .times. mouse microcell hybrids resistant to G418 were constructed between mouse hepatoma cells and human .times. mouse whole cell hybrids containing only intact human chromosome 5 and 22 with an integrated neor-gene. Among these, microcell hybrid BG15 produced four subclones, BG15-4, BG15-6, BG15-7 and BG15-9, which contained variously sized complements of human chromosome 5. BG15-6 contained an intact human chromosome 5, BG15-7 a deleted human chromosome 5 (5pter-q22) and PG15-4 and BG15-9 a translocation between parts of human chromosome 5 (pter-qter? and pter-q23, respectively) and a mouse chromosome. Southern DNA blot analysis showed that the human dihydrofolate reductase (DHFR) gene was present in all four subclones, whereas the human homolog of the v-fms gene was present in BG15-4 and 15-6, but absent from BG15-7 and 15-9. BG15-4, 15-6 and 15-9 were sensitive to diphtheria toxin, and only BG15-7 was resistant to the toxin. We used these microcell hybrids to restrict further the regional location of the gene for diphtheria toxin sensitivity to the q23 region of human chromosome 5.