Open AccessDeleteagene: a fast neutron deletion mutagenesis‐based gene knockout system for plants
Author(s) -
Li Xin,
Lassner Michael,
Zhang Yuelin
Publication year - 2002
Publication title -
comparative and functional genomics
Language(s) - English
Resource type - Journals
eISSN - 1532-6268
pISSN - 1531-6912
DOI - 10.1002/cfg.148
Subject(s) - mutant , mutagenesis , biology , genetics , gene , arabidopsis , gene knockout , homologous recombination , gene targeting , mutation
Deleteagene™ (Delete‐a‐gene) is a deletion‐based gene knockout systemfor plants. To obtain deletion mutants for a specific gene, random deletionlibraries created by fast neutron mutagenesis are screened by polymerase chainreaction (PCR) using primers flanking the target gene. By adjusting the PCRextension time to preferentially amplify the deletion alleles, deletion mutantscan be identified in pools of DNA samples with each sample representing morethan a thousand mutant lines. In Arabidopsis , knockout plants for greaterthan 80% of targeted genes have been obtained from a population of 51 840lines. A large number of deletion mutants have been identified and multipledeletion alleles are often recovered for targeted loci. In Arabidopsis ,the method is very useful for targeting small genes and can be used to finddeletion mutants mutating two or three tandem homologous genes. In addition,the method is demonstrated to be effective in rice as a deletion mutant fora rice gene was obtained with a similar approach. Because fast neutron mutagenesisis applicable to all plant genetic systems, Deleteagene™ has the potentialto enable reverse genetics for a wide range of plant species. Copyright ©2002 John Wiley & Sons, Ltd.