刺参PDRG基因全长克隆及其夏眠期间在消化道中的表达特征  

Cloning and characterization of PDRG gene from sea cucumber Apostichopus japonicus and the expression in intestine during aestivation

作　　者：王天明[1,2] 杨红生[1] 

机构地区：[1]中国科学院海洋研究所,海洋生态与环境科学重点实验室,山东青岛266071 [2]中国科学院研究生院,北京100049

出　　处：《海洋科学》2013年第12期1-9,共9页Marine Sciences

基　　金：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"

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。Sea cucumber （Apostichopus japonicus）undergoes an inactive phase “aestivation” as an important sur-vival strategy in high-temperature environments. During aestivation, the tissue of A. japonicus especially the intes-tine 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 func-tion in tissue regression during the aestivation of A. japonicus, the PDRG was cloned, characterized and quantified in this study. The full-length cDNA sequence （1122bp） of PDRG gene from A. japonicus 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 isoelec-tric point is 7.83. Sea cucumbers were induced to aestivation at temperature of 25℃ to investigate the transcrip-tional variation of PDRG during the intestine regression of aestivating A. japonicus. 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 expres-sion was observed at other sampling during the experiment. It was shown that the increase of PDRG gene expres-sion was closely associated with the regression of intestine during aestivating of A. japonicus. In summary, the re-sults 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.

关 键 词：刺参(Apostichopus japonicas) PDRG 夏眠 器官退化 实时定量PCR 

分 类 号：S96[农业科学—水产养殖] S917[农业科学—水产科学]