Hanaoka K, Hayasaka M, Uetsuki T, Fujisawa-Sehara A, Nabeshima Y
Division of Animal Models for Human Disease, National Institute of Neuroscience, Tokyo, Japan.
Differentiation. 1991 Dec;48(3):183-9. doi: 10.1111/j.1432-0436.1991.tb00256.x.
We have developed a method of marking of mouse cells by means of transfection of a foreign gene. The transgene chosen here was the plasmid pEF321CAT which contains the bacterial chloramphenicol acetyl transferase (CAT) gene linked to the promoter region of the human polypeptide chain elongation factor 1 alpha (hEF1 alpha) gene. Evaluation of the plasmid pEF321CAT as a cellular marker for mouse cells involved intensive examination of a transgenic mouse carrying pEF321CAT. The CAT gene was expressed in all tissues examined, demonstrating that the hEF1 alpha promoter was active in a wide range of mouse cells. The plasmid itself did not exert any harmful effect on the normal development of mice, and the CAT activity was immunohistologically detectable on sectioned tissues by the use of anti-CAT serum. When the plasmid was transferred into embryonal carcinoma (EC) cells and embryonic stem (ES) cells, the CAT gene was also found to be expressed constantly irrespective of their differentiation. These results demonstrated that the plasmid pEF321CAT can be used as a reliable and feasible cellular marker that would distinguish unequivocally the cells of each of genotype in chimeric tissues.
我们已经开发出一种通过转染外源基因来标记小鼠细胞的方法。这里选用的转基因是质粒pEF321CAT，它含有与人类多肽链延伸因子1α（hEF1α）基因启动子区域相连的细菌氯霉素乙酰转移酶（CAT）基因。将质粒pEF321CAT作为小鼠细胞的细胞标记进行评估，涉及对携带pEF321CAT的转基因小鼠进行深入检查。CAT基因在所有检测的组织中均有表达，表明hEF1α启动子在多种小鼠细胞中具有活性。该质粒本身对小鼠的正常发育未产生任何有害影响，并且通过使用抗CAT血清，在切片组织上可通过免疫组织化学方法检测到CAT活性。当该质粒转入胚胎癌细胞（EC）和胚胎干细胞（ES）时，发现CAT基因也持续表达，与它们的分化状态无关。这些结果表明，质粒pEF321CAT可作为一种可靠且可行的细胞标记，能够明确区分嵌合组织中每种基因型的细胞。
