Sorrell D A, Chrimes D, Dickinson J R, Rogers H J, Francis D
School of Biosciences, Cardiff University, PO Box 915, Cardiff, CF10 3TL, UK.
New Phytol. 2005 Feb;165(2):425-8. doi: 10.1111/j.1469-8137.2004.01288.x.
The putative mitotic inducer gene, Arath;CDC25 cloned in Arabidopsis thaliana, was screened for cell cycle function by overexpressing it in Schizosaccharomyces pombe (fission yeast). The expression pattern of Arath;CDC25 was also examined in different tissues of A. thaliana. Fission yeast was transformed with plasmids pREP1 and pREP81 with the Arath;CDC25 gene under the control of the thiamine-repressible nmt promoter. Using reverse transcription-polymerase chain reaction (RT-PCR), the expression of Arath;CDC25 was examined in seedlings, flower buds, mature leaves and stems of A. thaliana; actin (ACT2) was used as a control. In three independent transformants of fission yeast, cultured in the absence of thiamine (T), pREP1::Arath;CDC25 induced a highly significant reduction in mitotic cell length compared with wild type, pREP::Arath;CDC25 +T, and empty vector (pREP1 +/- T). The extent of cell shortening was greater using the stronger pREP1 compared with the weaker pREP81. However, Arath;CDC25 was expressed at low levels in all tissues examined. The data indicate that Arath;CDC25 can function as a mitotic accelerator in fission yeast. However, unlike other plant cell cycle genes, expression of Arath;CDC25 was not enhanced in rapidly dividing compared with non-proliferative Arabidopsis tissues.
通过在粟酒裂殖酵母（裂殖酵母）中过表达拟有丝分裂诱导基因At;CDC25（在拟南芥中克隆得到）来筛选其细胞周期功能。还检测了At;CDC25在拟南芥不同组织中的表达模式。用携带受硫胺素可抑制的nmt启动子控制的At;CDC25基因的质粒pREP1和pREP81转化裂殖酵母。使用逆转录-聚合酶链反应（RT-PCR）检测At;CDC25在拟南芥幼苗、花芽、成熟叶和茎中的表达；以肌动蛋白（ACT2）作为对照。在无硫胺素（T）条件下培养的三个独立的裂殖酵母转化体中，与野生型、pREP::At;CDC25 +T和空载体（pREP1 +/- T）相比，pREP1::At;CDC25诱导有丝分裂细胞长度显著缩短。与较弱的pREP81相比，使用较强的pREP1时细胞缩短程度更大。然而，At;CDC25在所检测的所有组织中表达水平都较低。数据表明At;CDC25在裂殖酵母中可作为有丝分裂促进剂发挥作用。然而，与其他植物细胞周期基因不同，与非增殖性拟南芥组织相比，At;CDC25在快速分裂组织中的表达并未增强。
