Molecular analyses of in vivo hprt mutations in human T-lymphocytes. I. Studies of low frequency 'spontaneous' mutants by Southern blots.
Molecular analyses of in vivo hprt mutations in human T-lymphocytes. I. Studies of low frequency 'spontaneous' mutants by Southern blots.
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人 T 淋巴细胞体内 hprt 突变的分子分析。
DOI：
10.1093/mutage/2.5.341
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发表时间：
1987
期刊：
影响因子：
2.7
通讯作者：
Albertini,RJ
中科院分区：
文献类型：
--
作者：
Nicklas,JA;Hunter,TC;Sullivan,LM;Berman,JK;O'Neill,JP;Albertini,RJ
从8名未暴露于诱变剂的成年男性中获得50个野生型和164个体内衍生的dhprt突变t细胞克隆，突变频率值在正常范围内（通常< 10 × 10−6），通过Southern blot分析来确定hprt基因总体结构改变的频率和程度。16个（9.8%）突变克隆出现了这种变化。病变部位和类型均不占优势。在研究可检测到的范围内，8个个体中恢复的突变体的总体结构改变的相对频率没有差异。用t细胞受体(TCR) β基因探针对214个克隆中的201个克隆的DNA进行了研究，作为体外克隆独立性的标记。一些克隆也用TCR γ基因探针进行了研究。发现94%的野生型和89%的突变体来源于独立的活体前体细胞。因此，研究中发现的大多数突变可能来自于单独的突变。对于具有正常突变频率的未诱变成人，因此，胞内突变频率是胞内突变频率的合理近似值，尽管我们在其他地方表明，对于突变频率严重升高的个体来说，这并不一定正确。
Fifty wild-type and 164in vivo-derivedhprtmutant T-cell clones obtained from eight non-mutagen-exposed adult males with mutant frequency values in the normal range (usually < 10 × 10−6) were studied by Southern blot analyses to determine frequency and extent of gross structural alterations in thehprtgene. Sixteen (9.8%) of the mutant clones showedhprtchanges. No site or type of lesion predominated. Relative frequencies of gross structural alterations in the recoveredhprtmutants did not differ among the eight individuals, within limits detectable by the study. DNA from 201 of these 214 clones was also studied with a T-cell receptor (TCR) β gene probe as a marker for independence ofin vivo-derived clones. Some clones were also studied with a TCR γ gene probe. Ninety-four percent of wild-type and 89% of thehprtmutants were found to originate from independentin vivoprecursors. Therefore, most of the recoveredhprtmutants in the study were presumably derived from separatein vivomutations. For non-mutagenized adults with normal mutant frequencies,in vivomutant frequencies are thus reasonable approximations ofin vivomutation frequencies, although elsewhere we show that this is not necessarily true for individuals with grossly elevated mutant frequencies.