中国卤虫滞育胚胎解除过程中hyou1基因的分子克

发布时间：2018-06-29 12:49

  本文选题：中国卤虫 + hyou1　； 参考：《辽宁师范大学》2016年硕士论文

【摘要】：中国卤虫(Artemia sinica)广泛分布于高盐水体中,遇到环境胁迫可以产生休眠卵,其胚胎滞育期新陈代谢减慢,包囊可以抵御低氧、高盐、寒冷等恶劣环境的影响。缺氧上调蛋白Hypoxia up-regulated 1,也被称为Orp150,其属于热休克蛋白家族的成员之一。Hypoxia up-regulated 1在低氧条件下的大鼠胶质细胞中首次被发现,积累在缺氧条件下的内质网内,被认为在蛋白质折叠和分泌中发挥重要作用。本研究采用RACE技术首次得到了中国卤虫Hypoxia up-regulated 1基因的cDNA全长As-hyou1,同时也对As-hyou1进行了生物信息学分析和分子进化等方面的分析。As-hyou1的基因全长为2952bp,As-hyou1基因包含了116bp的5’非编码区以及180bp的3’非编码区,其中2656bp为开放阅读框结构。As-hyou1基因所编码的HYOU1蛋白是一含有886个氨基酸残基的蛋白,具有典型的PTZ00009结构域,属于NBD糖激酶HSP70-actin超家族成员。理论分子量100.5KD,等电点为5.34。Signal P4.1检测到蛋白序列中含信号肽序列,故中国卤虫HYOU1为分泌性蛋白。利用原位杂交技术对As-hyou1的表达部位进行了分析,As-hyou1广泛分布于胚胎发育的各时期。卤虫虫体生长发育至已分化阶段,卤虫的头胸部、腹部、体表、附肢都会检测到阳性信号。采用荧光实时定量PCR,对As-hyou1的表达模式进行了研究,结果显示,As-hyou1在卤虫胚胎滞育刚刚结束阶段也就是滞育卵重新回到其正常发育进程时具有较高的表达(0h),在早期胚胎发育期逐渐下调(5h-15h),在虫体发育到无节幼体时期表现出上升趋势;在高盐刺激下As-hyou1的表达由50‰开始逐步下调;同时低温刺激下表达量呈下降趋势。研究初步证明,As-hyou1介导调节抑制由缺氧引起的细胞程序性死亡,并通过内质网促进翻译、转化和处理其他相关蛋白。本研究对进一步阐明As-hyou1在中国卤虫滞育解除、早期胚胎发育所起的作用,及对中国卤虫滞育发生与解除现象的分子机制的研究具有较为重要的理论意义。
[Abstract]:Chinese Artemia (Artemia sinica) is widely distributed in high salt water. When exposed to environmental stress, dormant eggs can be produced, the metabolism of embryo diapause is slow, and the cyst can resist the effects of hypoxia, high salt, cold and so on. Hypoxia up-regulated 1, also known as Orp150, is a member of the heat shock protein family. Hypoxia up-regulated 1 was first found in hypoxic rat glial cells and accumulated in the endoplasmic reticulum under hypoxia. It is thought to play an important role in protein folding and secretion. In this study, the full-length As-hyou1 cDNA of Chinese Artemia Hypoxia up-regulated 1 gene was obtained by race technique for the first time. Meanwhile, the As-hyou1 was analyzed in bioinformatics and molecular evolution. The gene length of As-hyou1 gene was 2952bpnAs-hyou1 gene containing the 5'of 116bp. The non-coding region and the 3'non-coding region of 180bp, The HYOU1 protein encoded by the open reading frame structure. As-hyou1 gene of 2656bp is a protein containing 886 amino acid residues. It has a typical PTZ00009 domain and belongs to the superfamily of NBD kinase HSP70-actin. The theoretical molecular weight was 100.5 KD, the isoelectric point was 5.34.Signal P4.1, the signal peptide sequence was detected in the protein sequence, so HYOU1 of Artemia chinensis was secreted protein. The expression site of As-hyou1 was analyzed by in situ hybridization technique. As-hyou1 was widely distributed in all stages of embryonic development. The positive signals were detected in the head, chest, abdomen, body surface and appendage of Artemia. The expression pattern of As-hyou1 was studied by real-time fluorescence quantitative PCR. The results showed that As-hyou1 had high expression (0h) at the end of diapause, which was the stage of diapause, and down-regulated (5h-15h) at the early stage of embryo development. The expression of As-hyou1 decreased gradually from 50 鈥，

本文编号：2082164