The use of resolvases T4 endonuclease VII and T7 endonuclease I in mutation detection

被引:40
|
作者
Babon, JJ
McKenzie, M
Cotton, RGH
机构
[1] Natl Inst Med Res, London NW7 1AA, England
[2] St Vincents Hosp, Mutat Res Ctr, Fitzroy, Vic 3065, Australia
关键词
mutation; mutation detection; polymorphism; resolvase; fluorescent; mismatch; hetero-duplex;
D O I
10.1385/MB:23:1:73
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mutation and polymorphism detection is of increasing importance in the field of molecular genetics. This is reflected by the plethora of chemical, enzymatic, and physically based methods of mutation detection. The ideal method would detect mutations in large fragments of DNA and position them to single base-pair (bp) accuracy. Few methods are able to quickly screen kilobase lengths of DNA and position the mutation at the same time. The Enzymatic Mismatch Cleavage (EMC) method of mutation detection is able to reliably detect nearly 100% of mutations in DNA fragments as large as 2 kb and position them to within 6 bp. This method exploits the activity of a resolvase enzyme from T4, T4 endonuclease VII, and, more recently, a second bacteriophage resolvase, T7 endonuclease I. The technique uses these enzymes to digest heteroduplex DNA formed by annealing wild-type and mutant DNA. Digestion fragments indicate the presence, and the position, of any mutations. This method is robust and reliable and much faster and cheaper than sequencing. These attributes have resulted in its increasing use in the field of mutation detection.
引用
收藏
页码:73 / 81
页数:9
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