Marchetti G, Evangelisti A, Bernardi F, Perrotta C, Conconi F, Del Senno L
Boll Soc Ital Biol Sper. 1983 Jun 30;59(6):758-64.
A library of genomic DNA was prepared from a patient with beta o Ferrara thalassaemia: random human DNA fragments (15 - 20 Kb) have been joined to phage lambda vectors and cloned has viable phage particles (4). 4x10(5) phages have been screened for their content in beta globin gene sequences, using a human beta cDNA plasmid (5) as hybridization probe. Five positive clones have been isolated and characterized by restriction endonuclease cleavage analysis and by the hybridization experiments. The results obtained allow the precise localization of the human fragments inside the beta like globin gene cluster (6). The comparison of the thalassaemic fragments with the normal DNA (6 - 7) shows two different restriction endonuclease sites, for Xba I and Eco RI respectively, downstream from the human beta globin gene.
从一名β型费拉拉地中海贫血患者制备了基因组DNA文库:随机的人类DNA片段(15 - 20 Kb)已连接到λ噬菌体载体上,并克隆出有活力的噬菌体颗粒(4)。使用人β cDNA质粒(5)作为杂交探针,对4×10⁵个噬菌体进行了β珠蛋白基因序列含量筛选。已分离出五个阳性克隆,并通过限制性内切酶切割分析和杂交实验进行了表征。所获得的结果允许将人类片段精确地定位在β类珠蛋白基因簇内(6)。地中海贫血片段与正常DNA(6 - 7)的比较显示,在人β珠蛋白基因下游分别有两个不同的限制性内切酶位点,分别对应Xba I和Eco RI。