农杆菌介导的拟南芥 C24 叶片转化的改进。

Improvements in the transformation of Arabidopsis thaliana C24 leaf-discs by Agrobacterium tumefaciens.

机构信息

Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333, AL Leiden, The Netherlands.

出版信息

Plant Cell Rep. 1996 Apr;15(8):572-7. doi: 10.1007/BF00232455.

DOI:10.1007/BF00232455

PMID:24178520

Abstract

We report here an efficient Arabidopsis leafdisc transformation protocol yielding an average transformation frequency of 1.6 transgenic shoots per leaf explant 4 weeks after the bacterial infection period. Subsequent cultivation in vitro is such that a high percentage (85-90%) of the primary transformants produces seeds with an average seed yield of 100-300 seeds per plant. This improved transformation protocol yields mainly (70%) transformants segregating for a single T-DNA locus of which 68% actually contain one T-DNA insert. The objective is to generate a pool of independent transformants harboring an activator T-DNA construct in a gene tagging approach to isolate genes involved in morphogenesis and auxin signal transduction.

摘要

我们在此报告了一种高效的拟南芥叶片圆盘转化方案，该方案在细菌感染期 4 周后，每个叶片外植体平均产生 1.6 个转基因芽。随后的体外培养使得较高比例（85-90%）的初级转化体产生种子，每个植物的平均种子产量为 100-300 粒。这种改进的转化方案主要产生（70%）分离单个 T-DNA 基因座的转化体，其中 68%实际上含有一个 T-DNA 插入。其目的是生成一个含有激活 T-DNA 构建体的独立转化体池，采用基因标记方法来分离参与形态发生和生长素信号转导的基因。