Magnusson K P, Minarovits J, Klein G, Wiman K G
Microbiology and Tumor Biology Center, Karolinska Institute, Stockholm, Sweden.
Gene. 1995 Sep 11;162(2):231-4. doi: 10.1016/0378-1119(95)00338-7.
The status of the p53 gene in SEWA-AS12-ADH (S-ADH) cells, a subline of the mouse sarcoma cell line SEWA, was examined. Immunoprecipitation with wild-type (wt) or mutant P53-specific monoclonal antibodies (mAb) showed that both wt and mutant P53 were produced. Sequence analysis of the p53 cDNA and genomic DNA revealed a single nucleotide (nt) substitution at a splice donor site at the beginning of intron 7. As a result of this mutation, an alternative splice site 15 nt further 3' in intron 7 is used. The P53 protein translated from this aberrantly spliced mRNA carries an Arg258-->Ser substitution, followed by an insertion of 5 extra amino acids. This is the first example of a splice-site mutation in the mouse p53 gene.
对小鼠肉瘤细胞系SEWA的亚系SEWA-AS12-ADH(S-ADH)细胞中的p53基因状态进行了检测。用野生型(wt)或突变型P53特异性单克隆抗体(mAb)进行免疫沉淀显示,wt和突变型P53均有产生。对p53 cDNA和基因组DNA的序列分析揭示,在内含子7起始处的一个剪接供体位点存在单核苷酸(nt)取代。由于该突变,使用了内含子7中3'端再下游15 nt处的一个替代剪接位点。从这种异常剪接的mRNA翻译出的P53蛋白携带Arg258→Ser取代,随后插入5个额外的氨基酸。这是小鼠p53基因中剪接位点突变的首个实例。