Developing the use of mismatch binding proteins for discovering rare somatic mutations
Publication in refereed journal


香港中文大学研究人员 ( 现职)

梁伟强教授 (内科及药物治疗学系)

包立怡教授 (药剂学院)

全文

数位物件识别号 (DOI) http://dx.doi.org/10.1016/j.mcp.2004.11.001

引用次数
Web of Sciencehttp://aims.cuhk.edu.hk/converis/portal/Publication/1WOS source URL

其它资讯

摘要A method for detecting rare, unknown. somatic mutations could allow presymptomatic cancer screening from human fluids. Immobilized mismatch binding protein can bind DNA heteroduplexes while allowing homoduplexes to be washed away, thus enriching for rare mutations. We examined the potential use. for mutation enrichment, of a fusion protein of maltose binding protein and the mismatch binding protein TaqMutS (MBP-MutS). Unlabeled and fluorescent-labeled oligonucleotides, either perfectly complementary or with single nucleotide mismatches or deletions. were combined to form homo- or heteroduplexes that were then mixed at low ratios of hetero- to homoduplexes and enriched for heteroduplexes. Enrichment was observed using a capillary DNA sequencer. A single base deletion oligonucleotide was enriched by a factor of 29 and a mismatch oligonucleotide was enriched by a factor of 2. N - http://aims.cuhk.edu.hk/converis/portal/Publication/1 oligonucleotide synthesis fragments were enriched more than were mismatches, suggesting that these deletion fragments may compete for MutS and impede enrichment of mismatches. Purification of oligonucleotides by high pressure liquid chromatography or polyacrylamide gel electrophoresis failed to remove n - http://aims.cuhk.edu.hk/converis/portal/Publication/1 fragments. thus overcoming this obstacle to enrichment of mismatch mutations may require alternative strategies, such as developing new purification methods or avoiding the use of synthetic oligonucleotides before enrichment. (c) 2004 Elsevier Ltd. All rights reserved.

着者Baum L, Ng A, Leung WK
期刊名称Molecular and Cellular Probes
出版年份2005
月份6
日期http://aims.cuhk.edu.hk/converis/portal/Publication/1
卷号http://aims.cuhk.edu.hk/converis/portal/Publication/19
期次3
出版社ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD
页次http://aims.cuhk.edu.hk/converis/portal/Publication/163 - http://aims.cuhk.edu.hk/converis/portal/Publication/168
国际标準期刊号0890-8508
语言英式英语

关键词enrichment; heteroduplex; MutS; oligonucleotide
Web of Science 学科类别Biochemical Research Methods; BIOCHEMICAL RESEARCH METHODS; Biochemistry & Molecular Biology; BIOCHEMISTRY & MOLECULAR BIOLOGY; Biotechnology & Applied Microbiology; BIOTECHNOLOGY & APPLIED MICROBIOLOGY; Cell Biology; CELL BIOLOGY