| Detection of mitochondrial DNA deletion by a modified PCR method |
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| 引用本文： | 汪振诚,王学敏,缪明永,章卫平,焦炳华,倪庆桂. Detection of mitochondrial DNA deletion by a modified PCR method[J]. 中国人民解放军军医大学学报, 2003, 18(3): 135-139 |
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| 作者姓名： | 汪振诚 王学敏 缪明永 章卫平 焦炳华 倪庆桂 |
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| 作者单位： | DepartmentofBiochemistryandMolecularBiology,CollegeofBasicMedicalSciences,SecondMilitaryMedicalUniversity,Shanghai200433,China |
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| 基金项目： | Supported by the National Natural Science Foundation of China(No.30171030) |
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| 摘 要： | Objective: To develop a simple and efficient method for detecting small populations of mitochondrial DNA deletion. Methods: Peripheral blood cell DNA was obtained from a victim who was aecidently exposed to a ^60Co radiation source 11 years ago. Using the DNA as template, PCR was performed to generate multiple products including true deletions and artifacts. The full length product was recovered and used as template of secondary PCR. The suspicious deletion product of mtDNA could be confirmed if it was only yielded by first PCR. Using either original primers or their nested primem, the suspicious deletion product was amplified and authenticated as true deletion product. The template was recovered and determined to be a deletion by sequencing directly. Results: A new mtDNA deletion, spanning 889 bp from nt11688 to nt12576, was detected in the peripheral blood cells of the victim. Conclusion: The new PCR-based method is more efficient in detecting small populations of mtDNA deletion than other routine methods. MtDNA deletion is found in the victim, suggesting there is relationship between the deletion and phenotypes of the disease.
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| 关 键 词： | 线粒体DNA 缺失突变 聚合酶链反应 ^60Co辐射 |
Detection of mitochondrial DNA deletion by a modified PCR method |
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| WANG Zhen-cheng,WANG Xue-min,MIAO Ming-yong,ZHANG Wei-ping,JIAO Bing-hua,NI Qing-gui. Detection of mitochondrial DNA deletion by a modified PCR method[J]. Journal of Medical Colleges of PLA(China), 2003, 18(3): 135-139 |
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| Authors: | WANG Zhen-cheng WANG Xue-min MIAO Ming-yong ZHANG Wei-ping JIAO Bing-hua NI Qing-gui |
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| Affiliation: | WANG Zhen-cheng,WANG Xue-min,MIAO Ming-yong,ZHANG Wei-ping,JIAO Bing-hua,NI Qing-guiDepartment of Biochemistry and Molecular Biology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433,China Hematological Department,Changhai Hospital,Second Military Medical University,Shanghai 200433,China |
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| Abstract: | Objective: To develop a simple and efficient method for detecting small populations of mitochondrial DNA deletion. Methods: Peripheral blood cell DNA was obtained from a victim who was accidently exposed to a 60Co radiation source 11 years ago. Using the DNA as template, PCR was performed to generate multiple products including true deletions and artifacts. The full length product was recovered and used as template of secondary PCR. The suspicious deletion product of mtDNA could be confirmed if it was only yielded by first PCR. Using either original primers or their nested primers, the suspicious deletion product was amplified and authenticated as true deletion product. The template was recovered and determined to be a deletion by sequencing directly. Results: A new mtDNA deletion, spanning 889 bp from nt11688 to nt12576, was detected in the peripheral blood cells of the Victim. Conclusion: The new PCR-based method is more efficient in detecting small populations of mtDNA deletion than other routine methods. MtDNA deletion is found in the victim, suggesting there is relationship between the deletion and phenotypes of the disease. |
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| Keywords: | PCR mitochondrial DNA deletion mutation 60Co radiation |
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