Krysan P J, Young J C, Tax F, Sussman M R
Cell and Molecular Biology Program and the Department of Horticulture, University of Wisconsin, Madison 53706, USA.
Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):8145-50. doi: 10.1073/pnas.93.15.8145.
The transferred DNA (T-DNA) of Agrobacterium tumefaciens serves as an insertional mutagen once integrated into a host plant's genome. As a means of facilitating reverse genetic analysis in Arabidopsis thaliana, we have developed a method that allows one to search for plants carrying F-DNA insertions within any sequenced Arabidopsis gene. Using PCR, we screened a collection of 9100 independent T-DNA-transformed Arabidopsis lines and found 17 T-DNA insertions within the 63 genes analyzed. The genes surveyed include members of various gene families involved in signal transduction and ion transport. As an example, data are shown for a T-DNA insertion that was found within CPK-9, a member of the gene family encoding calmodulin-domain protein kinases.
根癌农杆菌的转移DNA（T-DNA）一旦整合到宿主植物的基因组中，就会作为一种插入诱变剂。作为促进拟南芥反向遗传分析的一种手段，我们开发了一种方法，该方法能让人们在任何已测序的拟南芥基因中寻找携带F-DNA插入的植物。我们利用聚合酶链反应（PCR）筛选了9100个独立的T-DNA转化拟南芥株系，在分析的63个基因中发现了17个T-DNA插入。所检测的基因包括参与信号转导和离子转运的各种基因家族成员。例如，展示了在CPK-9（编码钙调蛋白结构域蛋白激酶的基因家族成员）中发现的一个T-DNA插入的数据。
