中国水仙NtSOC1基因克隆及亚细胞定位
发布时间：2018-10-25 15:47
【摘要】：以中国水仙‘金盏银台’为实验材料,采用RACE和RT-PCR技术获得1个与开花相关的转录因子(SOC1)的同源基因NtSOC1。NtSOC1的cDNA全长1 603bp,含有1个687bp开放阅读框,编码228个氨基酸。生物信息学分析表明,NtSOC1与单子叶植物的SOC1同源基因的氨基酸序列较为相似,且在C末端同样含有一个保守性很高的SOC1motif序列,说明NtSOC1是属于SOC1/TM3亚家族基因。荧光定量PCR分析显示,NtSOC1在花芽分化阶段的表达量随着花芽的分化而升高,花芽分化结束时减少,表明NtSOC1基因可能参与中国水仙的花芽分化。成功构建了NtSOC1基因表达载体pCAMBIA1302-NtSOC1,通过农杆菌转化洋葱表皮对编码蛋白进行亚细胞定位结果显示,NtSOC1基因编码蛋白定位于细胞核,符合转录因子的亚细胞定位特征。该实验结果为进一步研究NtSOC1基因的生物学功能奠定了基础。
[Abstract]:The cDNA of the homologous gene NtSOC1.NtSOC1 of anthesis related transcription factor (SOC1) was obtained by RACE and RT-PCR from Narcissus chinensis (Narcissus chinensis) as experimental material. The cDNA contained an open reading frame of 687bp encoding 228 amino acids. Bioinformatics analysis showed that the amino acid sequence of SOC1 homologous gene of NtSOC1 and Monocotyledonous plants was similar, and a highly conserved SOC1motif sequence was also found at the C-terminal, indicating that NtSOC1 belongs to the SOC1/TM3 subfamily gene. Fluorescence quantitative PCR analysis showed that the expression of NtSOC1 increased with flower bud differentiation and decreased at the end of flower bud differentiation, suggesting that NtSOC1 gene might be involved in flower bud differentiation of Narcissus chinensis. The NtSOC1 gene expression vector pCAMBIA1302-NtSOC1, was successfully constructed and transformed into onion epidermis by Agrobacterium tumefaciens for subcellular localization of the encoded protein. The results showed that the encoded protein of NtSOC1 gene was located in the nucleus and accorded with the subcellular localization characteristics of transcription factors. The results laid a foundation for further study of the biological function of NtSOC1 gene.
【作者单位】： 福建农林大学园艺学院;
【基金】：国家自然科学基金(31601783) 福建省自然科学基金(2014J01083)
【分类号】：Q943.2;S682.21
本文编号：2294147
[Abstract]:The cDNA of the homologous gene NtSOC1.NtSOC1 of anthesis related transcription factor (SOC1) was obtained by RACE and RT-PCR from Narcissus chinensis (Narcissus chinensis) as experimental material. The cDNA contained an open reading frame of 687bp encoding 228 amino acids. Bioinformatics analysis showed that the amino acid sequence of SOC1 homologous gene of NtSOC1 and Monocotyledonous plants was similar, and a highly conserved SOC1motif sequence was also found at the C-terminal, indicating that NtSOC1 belongs to the SOC1/TM3 subfamily gene. Fluorescence quantitative PCR analysis showed that the expression of NtSOC1 increased with flower bud differentiation and decreased at the end of flower bud differentiation, suggesting that NtSOC1 gene might be involved in flower bud differentiation of Narcissus chinensis. The NtSOC1 gene expression vector pCAMBIA1302-NtSOC1, was successfully constructed and transformed into onion epidermis by Agrobacterium tumefaciens for subcellular localization of the encoded protein. The results showed that the encoded protein of NtSOC1 gene was located in the nucleus and accorded with the subcellular localization characteristics of transcription factors. The results laid a foundation for further study of the biological function of NtSOC1 gene.
【作者单位】： 福建农林大学园艺学院;
【基金】：国家自然科学基金(31601783) 福建省自然科学基金(2014J01083)
【分类号】：Q943.2;S682.21
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