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题名: 秀丽隐杆线虫经福尔马林处理后上调表达基因的分析
作者: 陈星桃
答辩日期: 2008-06-02
导师: 王桂堂
授予单位: 中国科学院水生生物研究所
授予地点: 水生生物研究所
学位: 硕士
关键词: 秀丽隐杆线虫 ; 福尔马林 ; 抑制性差减杂交 ; 斑点杂交
其他题名: Analysis of up-regulated genes in Caenorhabditis elegans treated with formalin
摘要: 秀丽隐杆线虫(Caenorhabditis elegans)是一种重要的模式生物,目前已经被广泛应用到生物对各种驱虫剂抗性机制的研究中。福尔马林被普遍用于鱼类寄生虫病的防治中,但是,由于长期的应用,许多寄生虫对它产生了一定的抗药性。本研究将秀丽隐杆线虫接种到含800 ppm福尔马林的培养基中,传代培养一个月,应用抑制性差减杂交技术,筛选经福尔马林处理后表达上调的基因,共获得676个克隆。然后,对这些克隆运用斑点杂交技术做进一步筛选,得到表达具有显著差异的161个克隆,最后,对其进行了测序分析。 BLAST分析发现:(1)细胞凋亡相关基因的表达发生上调,这些基因编码包括线粒体呼吸链相关蛋白、含TPR序列的蛋白SGT-1、热休克蛋白、氧化应激相关蛋白、胞吞过程相关蛋白、DNA复制和修复相关蛋白以及其它一些重要的凋亡相关蛋白;(2)编码重要的转录调节因子和信号传导相关蛋白如转录激活因子FosB/c-Fos、G蛋白、细胞周期蛋白B、钙结合蛋白、核小体装配蛋白NAP-1等的基因的表达发生上调;(3)能量代谢和蛋白质、脂肪、氨基酸代谢途径中的有些基因的表达也发生上调。(4)除了以上已知功能的基因外,还有一些未命名蛋白质基因。 这些结果说明福尔马林对秀丽隐杆线虫的影响除了诱导细胞凋亡之外,还伴随着其它细胞代谢活动的改变。本研究为进一步研究生物体对福尔马林的抗性机制奠定了的基础。
英文摘要: Caenorhabditis elegans, as good model organism, has been used in the study of resistant mechanism against anthelmintics. On the other hand, formalin has been widely used in prevention and cure for fish parasites. However, because of long term application, many parasites generate resistance to formalin. In the present study, after the nematode C. elegans was serially cultivated in the media containing 800ppm formalin for one month, the up-regulation genes were screened by using the methods of suppression subtractive hybridization. Totally 676 gene clones were got. Then, we use dot blot hybridization to screening the positive clones and got 161 gene clones that showed evident expression discrepancy. At last, the clones were sequenced. BLAST analyses indicate that: (1) Apoptosis related genes’ expression has been up-regulated. These genes encoding proteins include mitochondrial respiratory chain related protein, protein SGT-1 containing TPR motif, heat shock protein, oxidative stress related protein, endocytosis related protein, DNA duplication and reparation related protein and other important apoptosis related protein. (2) Those genes coding important transcription regulation factor and signal transduction related protein, for example, transcriptional activation factor FosB/c-Fos, G protein, cyclin B, Ca-binding protein, and nucleosome assembly protein NAP-1, have been up-regulated expression. (3) Some genes participating in energy metabolism, protein metabolism, fat metabolism and amino acid metabolism have also been up-regulated. (4) In addition, some gene clones coding unnamed protein have been obtained. The results indicate that besides inducing apoptosis., formalin also changes Caenorhabditis elegans‘ cell metabolism This study establishes foundation for further understanding of the resistance mechanism of organism to formalin.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.ihb.ac.cn/handle/342005/12356
Appears in Collections:中科院水生所知识产出(2009年前)_学位论文

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Recommended Citation:
秀丽隐杆线虫经福尔马林处理后上调表达基因的分析.陈星桃[d].中国科学院水生生物研究所,2008.20-25
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