RESTRICTION ENDONUCLEASE ANALYSIS OF MITOCHONDRIAL DNA FROM HUMAN LUNG ADENOCARCINOMA CELL LINE SPC-A-1
Abstract
Objective: To understand the role of mitoehondrial DNA (mIDNA) in carcinogenesis.
Methods: single-step method was used to isolate the mtDNA from human lung adenocarcinoma cell line SPC-A-1. The mtDNA was analyzed by restriction fragment length polymorphism (RFLP) with 11 kinds of restriction endonuclease, which were Pvu II, Xho I, Pst I, EcoR I, BstE II, Hind IIl, Hpa I, Bcl I, EcoR V, Sea I and Xba L Restriction map of mtDNA from SPC-A-1 cell was obtained by the single and double-digestion method.
Results: It was found that no variation at 32 restrictionsites could be detected in the coding region of mtDNA from SPC-A-1 cell line. But a new site was found at nueleotide 16276 (EeoR V) within the noneoding region.
Conclusion: These results indicate that the primary. structure of gene coding region of mtDNA isolated from SPC-A-1 cell is highly stable. While the major variation of nucleotide is probably located in the noncoding region.
Methods: single-step method was used to isolate the mtDNA from human lung adenocarcinoma cell line SPC-A-1. The mtDNA was analyzed by restriction fragment length polymorphism (RFLP) with 11 kinds of restriction endonuclease, which were Pvu II, Xho I, Pst I, EcoR I, BstE II, Hind IIl, Hpa I, Bcl I, EcoR V, Sea I and Xba L Restriction map of mtDNA from SPC-A-1 cell was obtained by the single and double-digestion method.
Results: It was found that no variation at 32 restrictionsites could be detected in the coding region of mtDNA from SPC-A-1 cell line. But a new site was found at nueleotide 16276 (EeoR V) within the noneoding region.
Conclusion: These results indicate that the primary. structure of gene coding region of mtDNA isolated from SPC-A-1 cell is highly stable. While the major variation of nucleotide is probably located in the noncoding region.