| 摘要: |
| 夏眠是刺参的重要生理特征; 夏眠期间, 刺参体重明显减轻, 器官萎缩、退化, 其中消化道退化明显。PDRG(p53 and DNA damage-regulated gene)是近年来研究发现与细胞凋亡具有密切联系的基因,其表达可促进细胞凋亡。本研究利用SMART RACE 技术克隆获得刺参PDRG 基因的cDNA 全长序列,并以此为基础, 研究刺参夏眠期间PDRG 基因表达与消化道退化的相关性。结果显示, 刺参PDRG 基因cDNA 全长为1122bp, 包含127bp 的5’非翻译区(untranslated region, UTR), 581bp 的3’UTR 和414bp的开放阅读框(open reading frame, ORF); ORF 区编码137 个氨基酸, 推算的分子质量约为16.1kDu, 理论等电点为7.83。研究通过25℃温度诱导刺参进入夏眠, 利用实时定量PCR 方法, 定量检测刺参消化道PDRG 基因表达, 结果表明: 夏眠期间刺参消化道PDRG 基因mRNA 表达水平与对照组相比出现显著上升, 实验5d 时显著上调至约2.49 倍, 10d 时显著上调至约1.51 倍, 而在0、20、40 d 时未检测出显著变化; 与刺参消化道相对质量变化数据结合分析表明, 刺参夏眠期间消化道的PDRG 基因高表达与其萎缩、退化密切相关。本研究阐明刺参夏眠期间消化道组织退化过程中PDRG 基因表达特征, 证明刺参PDRG 基因表达与消化道退化具有相关性, 为进一步探讨PDRG 基因在动物器官退化过程中的功能提供参考依据。 |
| 关键词: 刺参(Apostichopus japonicas) PDRG 夏眠 器官退化 实时定量PCR |
| DOI： |
| 分类号: |
| 基金项目:国家“863”高技术研究发展计划项目(2006AA10A411); 山东省农业良种工程课题“速生抗病耐高温刺参良种选育” |
|
| Cloning and characterization of PDRG gene from sea cucumber Apostichopus japonicas and the expression in intestine during aestivation |
|
|
| Abstract: |
| Sea cucumber (Apostichopus japonicas)undergoes an inactive phase “aestivation” as an important survival strategy in high-temperature environments. During aestivation, the tissue of A. japonicasespecially the intestine was degenerated. PDRG (p53 and DNA damage-regulated gene) was a novel gene which was found to facilitate the UV-induced cell killing and be involved in apoptosis. To investigate the gene expression of PDRG and its function in tissue regression during the aestivation of A. japonicas, the PDRG was cloned, characterized and quantified in this study. The full-length cDNA sequence (1122bp) of PDRG gene from A. japonicas was cloned by SMART RACE. It consists of a 127 bp 5’ UTR (untranslated region), a 581 bp 3’ TUR and a 414 bp ORF (open reading frame). The deduced protein is composed of 127 amino acids with a molecular weight of 16.1 kDa, and its isoelectric point is 7.83. Sea cucumbers were induced to aestivation at temperature of 25℃ to investigate the transcriptional variation of PDRG during the intestine regression of aestivatingA. japonicas. The intestines were sampled, at 0, 5, 10, 20 and 40 d after the temperature reached 25℃, and then analyzed using real-time PCR for PDRG gene quantification. During the thermal inducing aestivation experiment, the mRNA level of PDRG was increased to 2.49 fold at 5 d sampling time and 1.51 fold at 10 d compared to the control. No significant variation of PDRG expression was observed at other sampling during the experiment. It was shown that the increase of PDRG gene expression was closely associated with the regression of intestine during aestivating of A. japonicas. In summary, the results of this study indicate that the PDRG has potential function in facilitating tissue regression during aestivation, which has provided a reference to explore the function of PDRG gene in animal organs degeneration. |
| Key words: Apostichopus japonicas (Selenka) PDRG aestivation tissue regression Real-time PCR |
