Okuda M, Minehata K, Setoguchi A, Watari T, Goitsuka R, Tsujimoto H, Hasegawa A
Department of Veterinary Internal Medicine, Faculty of Agriculture, University of Tokyo, Japan.
Leukemia. 1997 Apr;11 Suppl 3:372-5.
For investigation of the relation of cell cycle regulation with tumorigenesis in cats, we cloned feline p21WAF1 and p27Kip1 cDNAs and searched for their aberration in feline spontaneous leukemias and lymphomas. The feline p21WAF1 cDNA (pCFW.31) clone obtained from the PCR amplified product appeared to cover approximately 75% of the open reading frame, and showed 81.6% and 76.8% sequence similarities with those of human and mouse counterparts, respectively. The pHFK.5 clone isolated by plaque hybridization contained the whole open reading frame of cat p27Kip1 cDNA encoding 198 amino acids, showing 93.4% and 90.4% sequence similarities with those of human and mouse counterparts, respectively. Southern-blot analyses using these clones as probes did not show any deletion or rearrangement of both the p21WAF1 and p27Kip1 genes in 19 feline spontaneous cases of leukemias and lymphomas examined. RT-PCR/SSCP (single strand conformation polymorphism) analysis of p27Kip1 cDNA indicated that there was no mutation resulting in amino-acid substitution in 10 feline leukemia and lymphoma cases.
为研究猫细胞周期调控与肿瘤发生的关系，我们克隆了猫的p21WAF1和p27Kip1 cDNA，并在猫的自发性白血病和淋巴瘤中寻找它们的畸变情况。从PCR扩增产物中获得的猫p21WAF1 cDNA（pCFW.31）克隆似乎覆盖了约75%的开放阅读框，与人类和小鼠相应序列的相似性分别为81.6%和76.8%。通过噬菌斑杂交分离的pHFK.5克隆包含了编码198个氨基酸的猫p27Kip1 cDNA的完整开放阅读框，与人类和小鼠相应序列的相似性分别为93.4%和90.4%。以这些克隆为探针进行的Southern杂交分析显示，在所检测的19例猫自发性白血病和淋巴瘤病例中，p21WAF1和p27Kip1基因均未出现任何缺失或重排。对p27Kip1 cDNA进行的RT-PCR/SSCP（单链构象多态性）分析表明，在10例猫白血病和淋巴瘤病例中没有导致氨基酸替代的突变。
