应用RDPCR技术检测饮用水有机浓集物对大鼠p53基因的损伤作用

Detection of DNA Damage of p53 Induced by Organic Concentrate of Drinking Water Using Terminal Linker,Dependent PCR

目的运用依赖随机化末端连接物聚合酶链式反应(RDPCR)技术检测环境中混合污染物的DNA损伤作用。方法将水样有机浓集物腹腔注射染毒SD大鼠,提取肝、肺、肾及白细胞的DNA,运用RDPCR技术检测其对大鼠p53基因外显子7的DNA损伤作用。结果水样有机浓集物对大鼠的基因组DNA具有断裂作用;经RDPCR、杂交显色后在大鼠肝、肾组织中检测到了杂交条带,而血及肺组织中未检测到杂交条带。结论RDPCR技术可成功地运用于检测环境混合污染物。水样有机浓集物对大鼠p53基因外显子7具有DNA损伤作用,其主要靶器官为肝和肾。

Objective To detect DNA lesions induced by complex pollutants existing in environment using the technique of randomized terminal linker,dependent PCR (RDPCR). Methods SD rats were treated with organic concentrate by injection i.p., DNA were extracted from rat's liver, kidney, lung and leucocytes, then RDPCR was used to detect DNA damage of exon 7 of p53 gene. Results Organic concentrate of drinking water could make rat's genomic DNA broken. Hybridization bands were found in liver and kidney tissues after RDPCR, none in blood and lung tissues. Conclusion Organic concentrate of drinking water can cause the DNA lesions of exon 7 of p53 gene and the major target organs are liver and kidney. The technique of RDPCR can be used to detect DNA damage induced by complex environmental pollutants.