氯霉素抗性线粒体DNA向嵌合小鼠的转移。
Transfer of chloramphenicol-resistant mitochondrial DNA into the chimeric mouse.
作者信息
Levy S E, Waymire K G, Kim Y L, MacGregor G R, Wallace D C
机构信息
Center for Molecular Medicine, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
出版信息
Transgenic Res. 1999 Apr;8(2):137-45. doi: 10.1023/a:1008967412955.
The mitochondrial DNA (mtDNA) chloramphenicol (CAP)-resistance (CAPR) mutation has been introduced into the tissues of adult mice via female embryonic stem (ES) cells. The endogenous CAP-sensitive (CAPS) mtDNAs were eliminated by treatment of the ES cells with the lipophilic dye Rhodamine-6-G (R-6-G). The ES cells were then fused to enucleated cell cytoplasts prepared from the CAPR mouse cell line 501-1. This procedure converted the ES cell mtDNA from 100% wild-type to 100% mutant. The CAPR ES cells were then injected into blastocysts and viable chimeric mice were isolated. Molecular testing for the CAPR mutant mtDNAs revealed that the percentage of mutant mtDNAs varied from zero to approximately 50% in the tissues analyzed. The highest percentage of mutant mtDNA was found in the kidney in three of the chimeric animals tested. These data suggest that, with improved efficiency, it may be possible to transmit exogenous mtDNA mutants through the mouse germ-line.
线粒体DNA（mtDNA）氯霉素（CAP）抗性（CAPR）突变已通过雌性胚胎干细胞（ES细胞）导入成年小鼠组织中。通过用亲脂性染料罗丹明-6-G（R-6-G）处理ES细胞，消除了内源性氯霉素敏感（CAPS）mtDNA。然后将ES细胞与从CAPR小鼠细胞系501-1制备的去核细胞质体融合。此过程将ES细胞的mtDNA从100%野生型转化为100%突变型。然后将CAPR ES细胞注射到囊胚中，并分离出存活的嵌合小鼠。对CAPR突变mtDNA的分子检测显示，在所分析的组织中，突变mtDNA的百分比从零到约50%不等。在三只受试嵌合动物的肾脏中发现了最高百分比的突变mtDNA。这些数据表明，随着效率的提高，有可能通过小鼠种系传递外源性mtDNA突变体。
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