Nicklas J A, Hunter T C, O'Neill J P, Albertini R J
Genetics Laboratory, University of Vermont, Burlington 05401.
Mutat Res. 1989 Dec;215(2):147-60. doi: 10.1016/0027-5107(89)90178-4.
The hprt clonal assay detects mutations occurring in vivo in the hypoxanthine-guanine phosphoribosyltransferase (hprt) gene of human T-lymphocytes. Analysis of 94 wild-type and 326 hprt mutant clones from 3 normal males was performed using Southern blotting with hprt and T-cell receptor (TCR) gene probes. Gross structural alterations of the hprt gene occurred in approximately 14% of the in vivo derived mutants. Breakpoints were randomly distributed across the gene with one possible mutational "hot spot" observed. Most hprt mutants were independent as judged by TCR gene rearrangement patterns indicating that the measured hprt mutant frequency is a good measure of the actual hprt mutation frequency. However, sibling mutants (generally doublets and triplets except for one nonamer) were detected. Information on the timing in vivo of the hprt mutational events and the persistence in vivo of sibling mutants was also obtained.
次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶（hprt）克隆分析可检测人类T淋巴细胞次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶（hprt）基因在体内发生的突变。使用hprt和T细胞受体（TCR）基因探针进行Southern印迹分析，对来自3名正常男性的94个野生型和326个hprt突变克隆进行了分析。hprt基因的总体结构改变发生在约14%的体内衍生突变体中。断点在整个基因中随机分布，观察到一个可能的突变“热点”。根据TCR基因重排模式判断，大多数hprt突变体是独立的，这表明测得的hprt突变频率是实际hprt突变频率的良好指标。然而，检测到了同胞突变体（除一个九联体之外，通常为双联体和三联体）。还获得了关于hprt突变事件在体内发生的时间以及同胞突变体在体内持续存在的信息。
