[Experiment research on the location of DNA lesion in the nucleic acid sequence].
OBJECTIVE
To research on establishing a method that can detect the DNA lesion at the level of nucleic acid.


METHODS
The TK6 cell was cultured and treated, the genomic DNA was extracted, and the strand cleavage was induced by the endonuclease Hinf I. Then the genomic DNA was amplified by randomized terminal linker-dependent PCR (RDPCR), and hybridized by Southern hybridization with the single-stranded probes of the exon 2 of k-ras gene. The PCR products were sequenced.


RESULTS
The clear hybridized band from the products of RDPCR was seen at the expectant position cut by Hinf I. The sequencing analysis showed that the position of DNA lesion linked by the linker was just the restriction site of Hinf I.


CONCLUSIONS
It can be used to detect the DNA lesion at the level of nucleic acid sequence with the combination of sequence and RDPCR technologies.
Gao
Xu-Fang
XF
Department of Environmental Health, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Rao
Cao-Long
CL
Wen
Wei-Hua
WH
Heng
Zheng-Cang
ZC
D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004249:DNA Damage; D005091:Exons / Q000235:genetics; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D004247:DNA