变性高效液相色谱在大通量筛查基因突变中的应用

顾颖; 李延; 陈蔚丰; 尹大强; 赵庆顺

环境化学

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顾颖, 李延, 陈蔚丰, 尹大强, 赵庆顺. 变性高效液相色谱在大通量筛查基因突变中的应用[J]. 环境化学, 2006, 25(3): 336-339.

引用本文:

顾颖, 李延, 陈蔚丰, 尹大强, 赵庆顺. 变性高效液相色谱在大通量筛查基因突变中的应用[J]. 环境化学, 2006, 25(3): 336-339.

GU Ying, LI Yan, CHEN Wei-feng, YIN Da-qiang, ZHAO Qing-shun. DHPLC AS A HIGH-THROUGHOUT METHOD FOR MUTATION ANALYSIS[J]. Environmental Chemistry, 2006, 25(3): 336-339.

Citation:

GU Ying, LI Yan, CHEN Wei-feng, YIN Da-qiang, ZHAO Qing-shun. DHPLC AS A HIGH-THROUGHOUT METHOD FOR MUTATION ANALYSIS[J]. Environmental Chemistry, 2006, 25(3): 336-339.

变性高效液相色谱在大通量筛查基因突变中的应用

1.
南京大学环境学院, 污染控制与资源化国家重点实验室, 南京, 210093;
2.
南京大学模式动物研究所, 医药生物技术国家重点实验室, 南京, 210061
基金项目:
国家自然科学基金(20577022)资助项目

DHPLC AS A HIGH-THROUGHOUT METHOD FOR MUTATION ANALYSIS

1.
School of the Environment, State Key Laboratory of Pollution Control and Resource Reuse, Nanjing University, Nanjing, 210093, China;
2.
Model Animal Research Center, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210061
Fund Project:

摘要: 应用变性高效液相色谱(DHPLC)对已知序列的含外显子7的斑马鱼p53基因片段PCR产物进行突变检测,结果表明,通过克隆测序方法检测出的点突变,用DHPLC同样可以检出.且对该片段的最优化检测柱温为60℃,其特异性很好,敏感性大于95%.
- 变性高效液相色谱 /
- 斑马鱼 /
- p53基因 /
- 点突变
Abstract: In this paper,we reported the optimal temperature and sensitivity of denaturing high-performance liquid chromatography(DHPLC) for mutation detection in zebrafish p53 gene fragment containing exon7.Our(results) showed that two point mutated samples determined by sequencing were also detectable by DHPLC.The optimal temperature to detect the point mutation of this fragment is 60℃ and the sensitivity for detecting(molecules) with point mutation is more than 95%.The results indicated that DHPLC can be developed to(detect) the point mutation of zebrafish p53 gene.
- DHPLC /
- zebrafish /
- p53 gene /
- point mutation

[1]

Xiao W,Oefner P J,Denaturing High-Performance Liquid Chromatography:A Review.Human Mutation,2001,17:439-474
[2] Kristensen V N,Kelefiotis D,Kristensen T et al.,High-Throughput Methods for Detection of Genetic Variation.Biotechniques,2001,30(2):318-322
[3] Jones A C,Sampson J R,Cheadle J P,Low Level Mosaicism Detectable by DHPLC But Not by Direct Sequencing.Human Mutation,2001,17:233-234
[4] Choy Y S,Dabora S L,Hall F et al.,Superiority of Denaturing High Performance Liquid Chromatography Over Single-Stranded Conformation and Conformation-Sensitive Gel Electrophoresis for Mutation Detection in TSC2.Ann.Hum.Genet.,1999,63:383-391
[5] Gross E,Kiechle M,Arnold N,Mutation Analysis of P53 in Ovarian Tumors by DHPLC.J Biochem Biophys Methods,2001,47:73-81
[6] Jones A C,Austin J,Hansen N et al.,Optional Temperature Selection for Mutation Detection by Denaturing HPLC and Comparison to Single-Stranded Conformation Polymorphism and Heteroduplex Analysis.Clin.Chem.,1999,45:1133-1140
[7] Amold N,Gross E,Schwarz-Boeger U et al.,A Highly Sensitive,Fast and Economical Technique for Mutation Analysis in Hereditary Breast and Ovarian Cancers.Human Mutation,1999,14:333-339
[8] Kuklin A,Munson K,Gjerde D et al.,Detection of Single Nucleotide Polymorphisms with the WAVE DNA Fragment Analysis System.Genet.Testing,1997,1(3):201-206
[9] Skopek T R,Glaab W E,Monroe J J et al.,Analysis of Sequence Alterations in a Defined DNA Region:Comparison of Temperature-Modulated Heteroduplex Analysis and Denaturing Gradient Gel Electrophoresis.Mutat.Res.,1999,430(1):13-21
[10] Shen J,Wang R T,Xu X P,Screening Unknown SNPs by Denaturing High Performance Liquid Chromatography(DHPLC).Foreign Medical Sciences(Section Genetics),2001,24(6):341-344

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变性高效液相色谱在大通量筛查基因突变中的应用

1. 南京大学环境学院, 污染控制与资源化国家重点实验室, 南京, 210093;
2. 南京大学模式动物研究所, 医药生物技术国家重点实验室, 南京, 210061

基金项目:

国家自然科学基金(20577022)资助项目

关键词:

摘要: 应用变性高效液相色谱(DHPLC)对已知序列的含外显子7的斑马鱼p53基因片段PCR产物进行突变检测,结果表明,通过克隆测序方法检测出的点突变,用DHPLC同样可以检出.且对该片段的最优化检测柱温为60℃,其特异性很好,敏感性大于95%.

English Abstract

DHPLC AS A HIGH-THROUGHOUT METHOD FOR MUTATION ANALYSIS

1. School of the Environment, State Key Laboratory of Pollution Control and Resource Reuse, Nanjing University, Nanjing, 210093, China;
2. Model Animal Research Center, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210061

Fund Project:

Keywords:

DHPLC /
zebrafish /
p53 gene /
point mutation

Abstract: In this paper,we reported the optimal temperature and sensitivity of denaturing high-performance liquid chromatography(DHPLC) for mutation detection in zebrafish p53 gene fragment containing exon7.Our(results) showed that two point mutated samples determined by sequencing were also detectable by DHPLC.The optimal temperature to detect the point mutation of this fragment is 60℃ and the sensitivity for detecting(molecules) with point mutation is more than 95%.The results indicated that DHPLC can be developed to(detect) the point mutation of zebrafish p53 gene.

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变性高效液相色谱在大通量筛查基因突变中的应用

DHPLC AS A HIGH-THROUGHOUT METHOD FOR MUTATION ANALYSIS

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变性高效液相色谱在大通量筛查基因突变中的应用

English Abstract

DHPLC AS A HIGH-THROUGHOUT METHOD FOR MUTATION ANALYSIS

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