Nonreciprocal homologous recombination between Agrobacterium transferred DNA and a plant chromosomal locus.
Nonreciprocal homologous recombination between Agrobacterium transferred DNA and a plant chromosomal locus.
复制标题
农杆菌转移的 DNA 与植物染色体基因座之间的非互易同源重组。
DOI：
10.1073/pnas.90.15.7346
复制
发表时间：
1993
影响因子：
11.1
通讯作者：
P. Hooykaas
中科院分区：
文献类型：
--
作者：
R. Offringa;M. F. Franke;M. D. de Groot;P. van den Elzen;P. Hooykaas
在此之前，我们证明了在农杆菌介导的转化后烟草细胞中发生基因打靶。在这些实验中，通过与含有不同缺陷Kmr基因的引入转移DNA（T-DNA）修复构建体（pSDM 101）的同源重组来恢复存在于染色体位置的缺陷卡那霉素抗性（Kmr）基因。在这篇文章中，我们描述了基因打靶实验与相同的目标线，但使用改进的修复结构，pSDM 321。在用pSDM 321（系A）转化后获得的Kmr愈伤组织之一中，用PCR检测同源重组产物。进一步的分子分析显示，引入的T-DNA上存在的缺陷Kmr基因已经通过与靶基因座的同源重组而恢复。靶基因座保持不变，发现校正的T-DNA插入在同一染色体上，但不靠近靶基因座。本文介绍了植物中引入的DNA分子（在这种情况下，T-DNA）通过同源染色体位点转化的分子证据。
Previously, we demonstrated the occurrence of gene targeting in tobacco cells after Agrobacterium-mediated transformation. In these experiments a defective kanamycin resistance (Kmr) gene residing at a chromosomal location was restored via homologous recombination with an incoming transferred DNA (T-DNA) repair construct (pSDM101) containing a different defective Kmr gene. In this article we describe gene targeting experiments with the same target line, but using an improved repair construct, pSDM321. In one of the Kmr calli obtained after transformation with pSDM321 (line A) the product of homologous recombination was detected using PCR. Further molecular analysis revealed that the defective Kmr gene present on the incoming T-DNA had been restored via homologous recombination with the target locus. The target locus was left unchanged and the corrected T-DNA was found to be inserted on the same chromosome but not close to the target locus. This paper presents molecular evidence in plants for the conversion of an introduced DNA molecule (in this case, T-DNA) by a homologous chromosomal locus.