Orkin S H
J Biol Chem. 1978 Jan 10;253(1):12-5.
Double-stranded human globin DNA synthesized in vitro from sickle cell mRNA has been used as a substrate for a series of restriction endonucleases. The double-stranded DNA contained full length transcripts of the alpha- and beta- globin genes. Of the 10 enzymes tested, only 3 (Hpa I, Sal I, and Kpn I) failed to cleave either alpha- or beta-DNA; 2 (Eco RI and Bam HI) cleaved only beta-DNA; 3 (HindIII, Hpa II, and Hha I) cleaved only alpha-DNA; and 2 (Hae III and Alu I) cleaved both alpha- and beta-DNAs. The selective cleavage of human globin genes by restriction endonucleases should provide a strategy for the identification and purification of DNA fragments of genomic DNA containing globin genes plus their flanking sequences, simplify the preparation of pure, chain-specific globin probes, and permit the isolation of DNA probes for specific regions of the globin genes.
从镰状细胞信使核糖核酸体外合成的双链人珠蛋白DNA已被用作一系列限制性内切酶的底物。双链DNA包含α-和β-珠蛋白基因的全长转录本。在测试的10种酶中,只有3种(Hpa I、Sal I和Kpn I)未能切割α-或β-DNA;2种(Eco RI和Bam HI)仅切割β-DNA;3种(HindIII、Hpa II和Hha I)仅切割α-DNA;2种(Hae III和Alu I)切割α-和β-DNA。限制性内切酶对人珠蛋白基因的选择性切割应为鉴定和纯化包含珠蛋白基因及其侧翼序列的基因组DNA片段、简化制备纯的、链特异性珠蛋白探针以及分离珠蛋白基因特定区域的DNA探针提供一种策略。