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通过分析拟南芥、豌豆、番茄和水稻的COP1(constitutively photomorphogenic 1)的cDNA序列,运用RT-PCR和改进的基因组步行(genome walking)技术相结合的方法,首次从甘蓝型油菜中克隆到油菜BnCOP1编码区cDNA的全长序列,其全长2 034 bp,编码677个氨基酸.同源性分析表明,其编码的氨基酸序列与拟南芥的同源性高达94%.对BnCOP1编码序列(cDNA)演绎出的氨基酸序列分析表明,其编码的蛋白包含有N端的环形锌指结合域(ring finger zinc bindingdomain,RING)、中间的卷曲螺旋形结构域(coiled-coil domain,coiled-coil),7个C端的WD-40重复序列(WD-40 repeats,WD-40)的功能域.半定量RT-PCR和实时荧光定量PCR分析该基因在油菜中的表达模式,结果显示,BnCOP1在甘蓝型油菜的各个组织器官中均有表达,其中在花中的表达明显高于在根、叶、茎、果荚及子叶和胚轴中,暗示该蛋白可能与开花途径相关.过表达BnCOP1的转基因拟南芥植株在高度、主茎的直径和叶片大小上都呈现出比野生型弱小的表型,表明BnCOP1抑制了拟南芥光形态建成从而影响了植物的生长发育.
By analyzing the cDNA sequence of COP1 (constitutively photomorphogenic 1) in Arabidopsis, pea, tomato and rice and using the combination of RT-PCR and improved genome walking technology, The full-length cDNA of BnCOP1 coding region was 2 034 bp in length and encoded 677 amino acids.The homology analysis indicated that the deduced amino acid sequence of BnCOP1 shared 94% identity with Arabidopsis thaliana.BnCOP1 coding sequence (cDNA ) Deduced amino acid sequence analysis showed that the encoded protein contains the N-terminal ring finger zinc binding domain (RING), the middle of the coiled-coil domain (coiled-coil domain, coiled-coil), 7 (WD-40 repeats, WD-40) domains were detected by semi-quantitative RT-PCR and real-time fluorescence quantitative PCR.The results showed that BnCOP1 was expressed in Brassica napus The expression of BnCOP1 in flowers was significantly higher than that in roots, leaves, stems, pods, cotyledons and hypocotyls, suggesting that this protein may be involved in the flowering pathway.The transgenic Arabidopsis thaliana Mustard plant The results showed that BnCOP1 inhibited the light morphogenesis of Arabidopsis thaliana and thus affected the growth and development of plants, both in height, diameter of main stem and leaf size.