ISOLATION AND CHARACTERIZATION OF LINKED DELTA-GLOBIN AND BETA-GLOBIN GENES FROM A CLONED LIBRARY OF HUMAN DNA
ISOLATION AND CHARACTERIZATION OF LINKED DELTA-GLOBIN AND BETA-GLOBIN GENES FROM A CLONED LIBRARY OF HUMAN DNA
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DOI：
10.1016/0092-8674(78)90043-0
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发表时间：
1978-01-01
期刊：
影响因子：
64.5
通讯作者：
MANIATIS, T
中科院分区：
文献类型：
--
作者：
LAWN, RM;FRITSCH, EF;MANIATIS, T
构建大的随机人胚胎DNA片段的克隆文库，并筛选β-使用克隆的人β-珠蛋白序列珠蛋白c[互补]DNA质粒pJW 102（Wilson等，1978）作为杂交探针。获得两个独立的克隆，然后通过限制性内切酶切割分析，杂交实验和部分DNA测序进行表征。每一个克隆体都携带着成年的.和β-珠蛋白基因这2个基因被大约5.4个DNA内切酶（kb）分开，它们相对于转录方向的取向是5“-δ-。β-的3''.两个. delta-和β-珠蛋白基因含有位于氨基酸104（精氨酸）和105（亮氨酸）的密码子之间的大的非编码间插序列（分别为950和900 bp）。虽然两个基因编码区内的大的间插序列的位置是相同的，但两个非编码序列具有很小的序列同源性。第二个较小的插入序列，类似于在其他哺乳动物β-在人β-珠蛋白的5“末端附近检测到珠蛋白基因。珠蛋白基因这两个独立分离的β-珠蛋白克隆彼此的不同之处在于在δ-珠蛋白的大的间插序列内存在Pst I限制酶切割位点。其中一个克隆的珠蛋白基因。2个克隆中携带的人DNA显然来自2条同源染色体，这2条染色体对于Pst I限制酶识别序列是杂合的。
A cloned library of large, random embryonic human DNA fragments was constructed and screened for .beta.-globin sequences using the cloned human .beta.-globin c[complementary]DNA plasmid pJW102 (Wilson et al., 1978) as a hybridization probe. Two independent clones were obtained and then characterized by restriction endonuclease cleavage analysis, hybridization experiments and partial DNA sequencing. Each of the clones carries both the adult .delta.- and .beta.-globin genes. The 2 genes are separated by approximately 5.4 kilobases (kb) of DNA and their orientation with respect to the direction of transcription is 5''-.delta.-.beta.-3''. Both the .delta.-and .beta.-globin genes contain a large noncoding intervening sequence (950 and 900 bp, respectively) located between the codons for amino acids 104 (arginine) and 105 (leucine). Although the location of the large intervening sequence within the coding regions of the 2 genes is identical, the 2 noncoding sequences bear little sequence homology. A 2nd, smaller intervening sequence similar to that found in other mammalian .beta.-globin genes was detected near the 5'' end of the human .beta.-globin gene. The 2 independently isolated .beta.-globin clones differ from each other by the presence of a Pst I restriction enzyme cleavage site within the large, intervening sequence of the .delta.-globin gene of one of the clones. The human DNA carried in the 2 clones was apparently derived from 2 homologous chromosomes which were heterozygous for the Pst I restriction enzyme recognition sequence.