Site-specific cleavage of human chromosome 4 mediated by triple-helix formation.
Site-specific cleavage of human chromosome 4 mediated by triple-helix formation.
复制标题
由三螺旋形成介导的人类 4 号染色体的位点特异性切割。
DOI：
10.1126/science.1836279
复制
发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Dervan,PB
中科院分区：
文献类型：
--
作者：
Strobel,SA;Doucette-Stamm,LA;Riba,L;Housman,DE;Dervan,PB
从人类基因组中直接物理分离特定的DNA片段是人类遗传学的一个必要目标。为了测试三螺旋介导的酶切是否可以释放人类染色体的特定片段，我们选择了人类4号染色体的尖端作为靶点，因为它包含了亨廷顿病基因的整个候选区域。16碱基嘧啶寡脱氧核糖核苷酸能够在超过10千兆酶对的基因组DNA中定位16碱基对嘌呤目标位点，并以超过80%的产量介导该位点的精确酶切。识别基序具有足够的普遍性，大多数宇宙体应该包含一个可被三螺旋结构靶向的序列。这种方法可以方便地将人类染色体从正常和受影响的个体分离成大碱基大小的片段，并方便候选基因位点的分离。
Direct physical isolation of specific DNA segments from the human genome is a necessary goal in human genetics. For testing whether triple-helix mediated enzymatic cleavage can liberate a specific segment of a human chromosome, the tip of human chromosome 4, which contains the entire candidate region for the Huntington's disease gene, was chosen as a target. A 16-base pyrimidine oligodeoxyribonucleotide was able to locate a 16-base pair purine target site within more than 10 gigabase pairs of genomic DNA and mediate the exact enzymatic cleavage at that site in more than 80 percent yield. The recognition motif is sufficiently generalizable that most cosmids should contain a sequence targetable by triple-helix formation. This method may facilitate the orchestrated dissection of human chromosomes from normal and affected individuals into megabase sized fragments and facilitate the isolation of candidate gene loci.