Optimization of DNA shuffling for high fidelity recombination

被引：166

作者：

Zhao, HM ^{[1
]}

Arnold, FH ^{[1
]}

机构：

[1] CALTECH,DIV CHEM & CHEM ENGN 21041,PASADENA,CA 91125

来源：

NUCLEIC ACIDS RESEARCH | 1997年 / 25卷 / 06期

关键词：

D O I：

10.1093/nar/25.6.1307

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：

A convenient 'DNA shuffling' protocol for random recombination of homologous genes in vitro with a very low rate of associated point mutagenesis (0.05%) is described, In addition, the mutagenesis rate can be controlled over a wide range by the inclusion of Mn2+ or Mg2+ during DNase I digestion, by choice of DNA polymerase used during gene reassembly as well as how the genes are prepared for shuffling (PCR amplification versus restriction enzyme digestion of plasmid DNA). These protocols should be useful for in vitro protein evolution, for DNA based computing and for structure-function studies of evolutionarily related genes.

引用

页码：1307 / 1308

页数：2

共 11 条

[1] TUNING THE ACTIVITY OF AN ENZYME FOR UNUSUAL ENVIRONMENTS - SEQUENTIAL RANDOM MUTAGENESIS OF SUBTILISIN-E FOR CATALYSIS IN DIMETHYLFORMAMIDE [J].