Seiser C, Beck G, Wintersberger E
Institut für Molekularbiologie, Universität Wien, Austria.
FEBS Lett. 1990 Sep 17;270(1-2):123-6. doi: 10.1016/0014-5793(90)81249-n.
Aside of the gene coding for cytoplasmic thymidine kinase, the genome of mouse cells carries two pseudogenes. Both are inactive in situ. One of the pseudogenes is a processed pseudogene in which a two base pair deletion caused a shift of the reading frame and a shortening of the gene product from the 233 amino acids of thymidine kinase to 177 amino acids in the pseudogene product. We report here that introduction of this pseudogene into LTK- cells gave rise to cells with a thymidine kinase positive phenotype. The transformed cells carried multiple copies of the pseudogene the upstream region of which exhibited low but measurable promoter activity. Replacement of the upstream region of the pseudogene by a promoter of Simian virus 40 or of the mammary tumor virus resulted in high transfection efficiencies and in cell lines exhibiting high thymidine kinase activities.
除了编码细胞质胸苷激酶的基因外,小鼠细胞基因组还携带两个假基因。两者在原位均无活性。其中一个假基因是一个加工假基因,其中两个碱基对的缺失导致阅读框移位,基因产物从胸苷激酶的233个氨基酸缩短为假基因产物中的177个氨基酸。我们在此报告,将该假基因导入LTK-细胞可产生具有胸苷激酶阳性表型的细胞。转化细胞携带该假基因的多个拷贝,其上游区域表现出低但可测量的启动子活性。用猿猴病毒40或乳腺肿瘤病毒的启动子替换假基因的上游区域,可导致高转染效率,并产生具有高胸苷激酶活性的细胞系。