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1985年,Jeffreys等首次报道用人肌红蛋白基因内含子中一个33bp的重复序列片段探针揭露出人小卫星DNA片段的高度多态性,并称之为DNA“指纹”。此后,若干学者对此进行了研究,并己逐渐应用于法医学亲权鉴定和个体识别的案件,取得了可喜的进展。 目前,国内人基因组DNA重复序列克隆还不易获得,用人工合成还较昂贵。我们根据随机探针检测DNA限制性片段长度多态性时,对探针的选择不在于它的来源或它原来的功能,重要的是能检测出多态性的原则,以及鼠与人的髓鞘碱性蛋白(MBP)基因cDNA有90％以上同源序列的事实,选用rMBP。cDNA3′-端非表达区高度重复序列的0.81Kb片段(从质粒pMBP-1获得),以α_32p-dCTP经随机引物延伸标记法标记后作探针。人白细胞DNA分别用HinfⅠ和HaeⅢ酸消化,经0.8％琼脂糖凝
In 1985, for the first time, Jeffreys et al. Reported that a 33 bp repeat probe in the human myoglobin gene intron revealed a high degree of polymorphism in human satellite DNA fragments and was referred to as DNA “fingerprinting.” Since then, a number of scholars have studied this and gradually applied forensic paternity testing and individual identification cases, and made encouraging progress. At present, domestic human genomic DNA clones are not easy to obtain cloning, using synthetic is also more expensive. When we detect DNA-restricted fragment length polymorphisms based on random probes, the choice of probe is not about its origin or its original function, but importantly the principle of detecting polymorphisms and the relationship between murine and human marrow The fact that rodent basic protein (MBP) cDNA has over 90% homology with rMBP was used. The 0.81Kb fragment (obtained from the plasmid pMBP-1) of the highly repetitive sequence of the non-expression region of the 3’-end of the cDNA was labeled with an a-32p-dCTP by a random primer extension labeling method. Human leukocyte DNA was digested with Hinf I and HaeIII acids, respectively, after 0.8% agarose