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本文介绍了人体淋巴细胞在体内突变和体外突变形成的测定和研究的详细方法。该方法包括制备含有白细胞介素-2的条件培养基,计数突变克隆,体外突变形成和进一步研究克隆的扩增问题。测定人体淋巴细胞突变而进行的克隆技术,为测定和研究人体体内突变以及研究个体细胞突变形成的敏感性提供了一种新的方法。突变可用测定对硫代鸟嘌呤有抗性的淋巴细胞克隆的频数来计数。该频数可用次黄嘌呤核糖基转移酶(HPRT)位点上所出现的突变来显示,其证据是该细胞失去了渗入放射性同位素标记的次黄嘌呤的能力,缺乏
This article describes a detailed method for the determination and study of human lymphocytes in vivo and in vitro. The method includes preparing a conditioned medium containing interleukin-2, counting mutant clones, mutating in vitro and further studying the amplification of the clones. The cloning technique for determining human lymphocyte mutations provides a new method for determining and investigating human in vivo mutations as well as investigating the susceptibility of individual cells to mutation formation. Mutations can be counted by determining the frequency of thioguanine-resistant lymphocyte clones. This frequency is shown by the presence of mutations in the hypoxanthine ribosyltransferase (HPRT) site, evidenced by the cell’s loss of ability to penetrate the radioisotope-labeled hypoxanthine, a deficiency