Conner B J, Reyes A A, Morin C, Itakura K, Teplitz R L, Wallace R B
Proc Natl Acad Sci U S A. 1983 Jan;80(1):278-82. doi: 10.1073/pnas.80.1.278.
Two 19-base-long oligonucleotides were synthesized, one complementary to the normal human beta-globin gene (beta A) and one complementary to the sickle cell beta-globin gene (beta S). The nonadecanucleotides were radioactively labeled and used as probes in DNA hybridization. Under appropriate hybridization conditions, these probes can be used to distinguish the beta A gene from the beta S allele. The DNA from individuals homozygous for the normal beta-globin gene (beta A beta A) only hybridized with the beta A specific probe; the DNA from those homozygous for the sickle cell beta-globin gene (beta S beta S) only hybridized with the beta S specific probe. The DNA from heterozygous individuals (beta A beta S) hybridized with both probes. This allele-specific hybridization behavior of oligonucleotides provides a general method for diagnosis of any genetic disease which involves a point mutation in the DNA sequence of a single-copy gene.
合成了两条19个碱基长的寡核苷酸,一条与正常人β-珠蛋白基因(βA)互补,另一条与镰状细胞β-珠蛋白基因(βS)互补。这些十九聚体寡核苷酸用放射性标记,并用作DNA杂交中的探针。在适当的杂交条件下,这些探针可用于区分βA基因和βS等位基因。纯合正常β-珠蛋白基因(βAβA)个体的DNA仅与βA特异性探针杂交;纯合镰状细胞β-珠蛋白基因(βSβS)个体的DNA仅与βS特异性探针杂交。杂合个体(βAβS)的DNA与两种探针都杂交。寡核苷酸的这种等位基因特异性杂交行为为诊断任何涉及单拷贝基因DNA序列点突变的遗传疾病提供了一种通用方法。