Churchill M E, Gemmell M A, Woloschak G E
Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, Illinois 60439-4833.
Radiat Res. 1994 Mar;137(3):310-6.
A polymerase chain reaction (PCR) technique has been developed to detect deletions in the mouse retinoblastoma gene using histological sections from radiation-induced and spontaneous tumors as the DNA source. Six mouse Rb gene exon fragments were amplified in a 40-cycle, 3-temperature PCR protocol. The absence of any of these fragments relative to control PCR products on a Southern blot indicated a deletion of that portion of the mouse Rb gene. Tumors chosen for analysis were lung adenocarcinomas that were judged to be the cause of death. Spontaneous tumors as well as those from irradiated mice (5.69 Gy 60Co gamma rays or 0.6 Gy JANUS neutrons, which have been found to have approximately equal radiobiological effectiveness) were analyzed for mouse Rb deletions. Tumors in 6 neutron-irradiated mice had no mouse Rb deletions. However, 1 of 6 tumors from gamma-irradiated mice (17%) and 6 of 18 spontaneous tumors from unirradiated mice (33%) showed a deletion in one or both mouse Rb alleles. All deletions detected were in the 5' region of the mouse Rb gene.
已开发出一种聚合酶链反应（PCR）技术，以辐射诱导的肿瘤和自发肿瘤的组织切片作为DNA来源，检测小鼠视网膜母细胞瘤基因中的缺失。在一个40循环、三温度的PCR方案中扩增了六个小鼠Rb基因外显子片段。在Southern印迹上，相对于对照PCR产物，这些片段中任何一个的缺失表明小鼠Rb基因的该部分发生了缺失。选择用于分析的肿瘤是被判定为死亡原因的肺腺癌。分析了自发肿瘤以及来自受辐照小鼠（5.69 Gy 60Coγ射线或0.6 Gy JANUS中子，已发现它们具有大致相等的放射生物学效应）的肿瘤中的小鼠Rb缺失情况。6只经中子辐照的小鼠的肿瘤未发生小鼠Rb缺失。然而，6只经γ射线辐照的小鼠中的1只肿瘤（17%）以及18只未受辐照的小鼠中的6只自发肿瘤（33%）在一个或两个小鼠Rb等位基因中显示出缺失。检测到的所有缺失均位于小鼠Rb基因的5'区域。
