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目的 建立一个用于临床标本的体外非同源末端连接的检测体系。方法 从白血病细胞株和正常骨髓及外周血细胞中提取的核蛋白在体外与线性质粒pUC18DNA一起作用,通过琼脂糖凝胶电泳分离和SYBRgreenⅠ染色观察其连接能力。结果 白血病细胞株NB4、U937、K5 6 2、SHI 1、Jurkat的连接能力为10 .6 %～32 .9% ,正常骨髓或外周血细胞的连接能力为5 .2 %～2 0 .0 %。结论 无细胞的非同源末端连接检测体系的建立是研究人类白血病细胞DNA修复机制的实验基础。
Objective To establish a detection system for in vitro non-homologous end-junctions of clinical specimens. Methods The nucleoprotein extracted from leukemia cell lines and normal bone marrow and peripheral blood cells was used in vitro to act on the linear plasmid pUC18DNA. The binding capacity was determined by agarose gel electrophoresis and SYBRgreenⅠ staining. Results The leukemia cell lines NB4, U937, K5 6 2, SHI 1 and Jurkat had a ligation capacity of 10.6% -32.9%, and the normal bone marrow or peripheral blood cells had a ligation capacity of 5.2% -2.0%. Conclusion The establishment of cell-free non-homologous end-joining assay system is the experimental basis for studying DNA repair mechanism of human leukemia cells.