Site‐specific integration of DNA into wild‐type and mutant <i>lox</i> sites placed in the plant genome

DOI Web Site Web Site 被引用文献19件

書誌事項

公開日
1995-04
権利情報
  • http://onlinelibrary.wiley.com/termsAndConditions#vor
DOI
  • 10.1046/j.1365-313x.1995.7040649.x
公開者
Wiley

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<jats:title>Summary</jats:title><jats:p>The bacteriophage P1 Cre—<jats:italic>lox</jats:italic> site‐specific recombination system has been used to integrate DNA specifically at <jats:italic>lox</jats:italic> sites previously placed in the tobacco genome. As integrated molecules flanked by wild‐type <jats:italic>lox</jats:italic> sites can readily excise in the presence of Cre recombinase, screening for mutant <jats:italic>lox</jats:italic> sites that can resist excisional recombination was performed. In gene integration experiments, wild‐type and mutant <jats:italic>lox</jats:italic> sites were used in conjunction with two strategies for abolishing post‐integration Cre activity: (i) promoter displacement of a <jats:italic>cre</jats:italic>‐expression construct present in the target genome; and (ii) transient expression of <jats:italic>cre</jats:italic>. When the promoter displacement strategy was used, integrant plants were recovered after transformation with constructs containing mutant <jats:italic>lox</jats:italic> sequences, but not with constructs containing wild‐type <jats:italic>lox</jats:italic> sites. When <jats:italic>cre</jats:italic> was transiently expressed, integrant plants were obtained after transformation with either mutant or wild‐type <jats:italic>lox</jats:italic> sites. DNA rearrangements at the target locus were less frequent when mutant <jats:italic>lox</jats:italic> sites were used. DNA integration at the genomic <jats:italic>lox</jats:italic> site was usually without additional insertions in the genome. Thus, the Cre—<jats:italic>lox</jats:italic> site‐specific recombination system is useful for the single‐copy integration of DNA into a chromosomal <jats:italic>lox</jats:italic> site.</jats:p>

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