Establishment of SV40-tsA58 transgenic rats as a source of conditionally immortalized cell lines.
Establishment of SV40-tsA58 transgenic rats as a source of conditionally immortalized cell lines.
复制标题
建立 SV40-tsA58 转基因大鼠作为条件永生化细胞系的来源。
作者：
R. Takahashi;M. Hirabayashi;N. Yanai;M. Obinata;M. Ueda
为了分离具有分化功能的多种大鼠细胞系，我们建立了在SV 40大T抗原本身控制下表达猿猴病毒40（SV 40）tsA 58突变体的温度敏感性大T抗原的转基因大鼠系。将该基因显微注射到564个Wistar大鼠卵细胞中，获得了23个独立的转基因候选体。十只幼鼠在断奶前死亡，八只转基因大鼠不能将转基因传递给后代。最后，建立了5个转基因大鼠品系。虽然一个品系（#1511-6）的繁殖力低，但其他四个品系的繁殖力正常。四个品系中的三个（#1507-2、#1509-7、#1519-8）表现正常，但另一个品系在3周龄时在脑和皮下组织中有肿瘤（#1511-6），在18至19周龄时在肾和皮下组织中有肿瘤（#1507-5）。从品系#1507-5和#1519-8的转基因胎仔制备的成纤维细胞表达转基因并表现出温度依赖性生长。通过转基因后代的同胞交配成功地产生了纯合的两个品系（#1507-5和#1519-8）。这些转基因大鼠系已经繁殖了许多代，并且已经被确立为新的条件永生化细胞系的现成来源。
To isolate a variety of rat cell lines with differentiated functions, we established transgenic rat lines expressing the temperature-sensitive large T-antigen of simian virus 40 (SV40) tsA58 mutant under the control of the SV40 large T-antigen itself. We microinjected the DNA into 564 eggs of Wistar rat and 23 independent transgenic candidates were obtained. Ten pups died before weaning and eight transgenic rats could not transmit the transgene to the progeny. Finally, five lines of the transgenic rat were established. Although one line (#1511-6) had low reproductivity, the other four lines reproduced normally. Three out of the four lines (#1507-2, #1509-7, #1519-8) appeared normal but the other line had tumors in the brain and subcutaneous tissue at 3 weeks of age (#1511-6), and in the kidneys and subcutaneous tissue at 18 to 19-weeks of age (#1507-5). Fibroblast cells prepared from transgenic fetuses of lines #1507-5 and #1519-8 expressed the transgene and exhibited temperature-dependent growth. Both of the lines (#1507-5 and #1519-8) were successfully generated to be homozygous by sibling mating of transgenic offspring. These transgenic rat lines have bred through many generations and have been established to be a ready source of novel conditionally immortalized cell lines.