Characterization of genes involved in regulation of differentiation using mutant plant cell cultured lines by insertion of transposable elements

通过插入转座元件，使用突变植物细胞培养系表征参与分化调节的基因

基本信息

批准号：
09440264
负责人：
OZEKI Yoshihiro
金额：
$ 8.64万
依托单位：
Tokyo University of Agriculture and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09440264/
关键词：
Daucus carota plant cell culture culture variation transposable element secondary metabolism anthocyanin

项目摘要

An En/Spm transposable elements of Tdc1 was found in the promoter region of phenylalanine ammonia-lyase gene in carrot suspension cultured cells. Tdc1 had only one long open reading frame corresponding to one transposase, but the other transposase, which is necessary for excision and insertion of Tdc1, was partially encoded in Tdc1, suggesting that Tdc1 should be a non-autonomous transposable element Southern analysis indicates that more than 20 copies of Tdc1-like elements may be in the carrot genome. The other elements of Tdc family were obtained using PCR and the nucleotide sequences were analyzed. Three sub-family of Tdc, Tdc-A, Tdc-B and TdcR, were found in carrot genome. Tdc-A1 encoded two transposases which were partially similar to maize En/Spm transposable element, suggesting that Tdc-A1 might be the autonomous element of Tdc family. The other Variants of Tdc-A showed the internal deletions without changes in nucleotide sequences. These results suggest that Tdc-A actively transposed accompanied with deletion during subculturing of carrot cells, which results in culture variation. Tdc-B and Tdc-R were deletion variants of Tdc-A.Their nucleotide sequences were changed in comparison with Tdc-A.They might transpose before the establishment of carrot cell culture.

在胡萝卜悬浮培养细胞中苯丙氨酸解氨酶基因启动子区发现Tdc1的En/Spm转座元件。 Tdc1只有一个长开放阅读框对应于一个转座酶，但Tdc1切除和插入所必需的另一种转座酶部分编码在Tdc1中，表明Tdc1应该是一种非自主转座元件Southern分析表明，更多胡萝卜基因组中可能存在超过 20 个 Tdc1 样元件拷贝。使用PCR获得Tdc家族的其他元件并分析核苷酸序列。在胡萝卜基因组中发现了Tdc的三个亚家族：Tdc-A、Tdc-B和TdcR。 Tdc-A1编码两个与玉米En/Spm转座元件部分相似的转座酶，表明Tdc-A1可能是Tdc家族的自主元件。 Tdc-A 的其他变体显示出内部缺失，但核苷酸序列没有变化。这些结果表明，在胡萝卜细胞传代培养过程中，Tdc-A 主动转座并伴有缺失，从而导致培养物变异。 Tdc-B和Tdc-R是Tdc-A的缺失变体，与Tdc-A相比，它们的核苷酸序列发生了变化。它们可能在胡萝卜细胞培养物建立之前发生了转座。