Smith-Sørensen B, Gebhardt M C, Kloen P, McIntyre J, Aguilar F, Cerutti P, Børresen A L
Department of Genetics, Norwegian Radium Hospital, Montebello, Oslo.
Hum Mutat. 1993;2(4):274-85. doi: 10.1002/humu.1380020407.
We have previously developed conditions to screen for TP53 point mutations inside the conserved domains II-V of the gene by using constant denaturant gel electrophoresis (CDGE). The present study reports conditions for screening more of the codons in the frequently mutated region exon 5-8 and for detecting mutations in sequences encoding functional domains in the N- and C-terminal part of the protein. The ability of the CDGE technique to detect mutations was studied using controls with known sequence deviations. The resolution power of the technique to separate different types of mutations was tested by using seven different single base pair mutants all residing in a stretch of four base pairs. All mutants were separated from the wild type. The established CDGE screening strategy was then used to look for mutations in DNA from 28 osteosarcomas. Six (21.5%) of the samples were shown to have a TP53 mutation, and the exact characterization was performed by direct sequencing. All of these were within the frequently reported mutated region exon 5-8.
我们之前已开发出相关条件，通过使用恒定变性剂凝胶电泳（CDGE）来筛选基因保守结构域II-V内的TP53点突变。本研究报告了用于筛选更多常见突变区域外显子5-8中密码子以及检测蛋白质N端和C端功能结构域编码序列中突变的条件。利用已知序列偏差的对照研究了CDGE技术检测突变的能力。通过使用均位于一段四个碱基对中的七个不同单碱基对突变体，测试了该技术分离不同类型突变的分辨能力。所有突变体均与野生型分离。然后使用既定的CDGE筛选策略来寻找28例骨肉瘤DNA中的突变。其中六个（21.5%）样本显示存在TP53突变，并通过直接测序进行了精确表征。所有这些突变均在经常报道的突变区域外显子5-8内。
