首页> 外文期刊>Fungal Genetics Newsletter >Split-Marker Recombination for Efficient Targeted Deletion of Fungal Genes
【24h】

Split-Marker Recombination for Efficient Targeted Deletion of Fungal Genes

机译:高效的真菌基因靶向删除的拆分标记重组。

获取原文
获取原文并翻译 | 示例
           

摘要

A commonly used method for fungal gene deletion is introduction of linear DNA consisting of a selectable marker gene flanked on both sides by short stretches of DNA that target a gene of interest (Wirsel et al 1996 Curr. Genet 29:241-249). Gene deletion in Cochliobolus heterostrophus and Gibberella zeae occurs efficiently with this approach. To facilitate deletion construct synthesis, we have applied the "split-marker" deletion strategy previously developed for Saccharomyces cerevisiae (Fairhead et al. 1996 Yeast 12:1439-57; Fairhead etal. 1998 Gene 223:33-46). Here, we describe both fusion PCR-based and plasmid-based deletion methods using this strategy with PEG-mediated protoplast transformation (Turgeon et al, 1985 Mol. Gen. Genet. 201:450-453). These methods are predicted to work well with any transformable fungus that undergoes homologous recombination between chromosomal and introduced DNA sequences.
机译:真菌基因缺失的常用方法是引入线性DNA,该线性DNA由选择性标记基因组成,该选择性标记基因在两侧两侧是靶向目的基因的短段DNA(Wirsel等,1996 Curr.Genet 29:241-249)。用这种方法可以有效地发生异链球菌和玉米赤霉菌中的基因缺失。为了促进缺失构建体的合成,我们应用了先前为酿酒酵母开发的“分裂标记”缺失策略(Fairhead等,1996 Yeast 12:1439-57; Fairhead等,1998 Gene 223:33-46)。在这里,我们描述了使用这种策略与PEG介导的原生质体转化的基于融合PCR的和基于质粒的缺失方法(Turgeon等,1985 Mol.Gen.Genet.201:450-453)。预计这些方法对于在染色体和导入的DNA序列之间进行同源重组的任何可转化真菌均适用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号