Roseman B, Lough J, Houkom E, Herman T
Biochem Biophys Res Commun. 1986 May 29;137(1):474-9. doi: 10.1016/0006-291x(86)91234-9.
Chicken erythrocyte nuclei were nick translated with the chemically cleavable biotinylated nucleotide, Bio-12-SS-dUTP. DNA was purified, digested with restriction endonucleases, and applied to an avidin-agarose affinity column. Seventy percent of the nick translated DNA bound to the column. This DNA was recovered from the column by chemical cleavage of the linker arm joining biotin to the DNA. Dot hybridization analysis of this DNA revealed a significant enrichment of the alpha-D-globin gene. This result suggests an approach to isolate transcriptionally active genes.
用化学可裂解的生物素化核苷酸Bio-12-SS-dUTP对鸡红细胞核进行缺口平移。纯化DNA，用限制性内切酶消化，然后应用于抗生物素蛋白-琼脂糖亲和柱。70%的缺口平移DNA与柱结合。通过化学裂解连接生物素与DNA的连接臂，从柱上回收该DNA。对该DNA的点杂交分析显示α-D-珠蛋白基因有显著富集。这一结果提示了一种分离转录活性基因的方法。
