文章摘要
陈思弘,周志刚.缺刻缘绿藻转录组测序及脂质代谢相关基因注释[J].上海海洋大学学报,2012,21(5):662-670
缺刻缘绿藻转录组测序及脂质代谢相关基因注释
The transcriptome pyrosequencing and gene function annotation of the green microalga Myrmecia incisa
  
DOI:
中文关键词: 缺刻缘绿藻  转录组  焦磷酸测序  脂质代谢途径
英文关键词: Myrmecia incisa  transcriptome  pyrosequencing  lipid metabolism
基金项目:国家自然科学基金(30972243、31172389);国家海洋局海洋可再生能源专项基金项目(SHME2011SW02);上海市教育委员会海洋生物学重点学科(J50701)
作者单位
陈思弘 上海海洋大学 水产与生命学院 
周志刚 上海海洋大学 水产与生命学院 
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中文摘要:
      为了能深入地了解缺刻缘绿藻花生四烯酸(ArA)和脂质的代谢过程,利用Roche 454 GS FLX测序仪对该藻转录组进行高通量的焦磷酸测序。得到高质量读序(read)382 468条,占原始读序的97.14%,平均每条读序长322 bp,总大小达123 Mb。经CAP3软件拼接得到22 714条重叠群、25 621条singleton。将这些序列与公共数据库进行同源性搜索、比较、基因功能注释和分类。基于转录组中所注释的基因构建缺刻缘绿藻脂质代谢途径:脂肪酸是在叶绿体内从头合成,然后游离脂肪酸进入胞质,由内质网进行三酰甘油的合成,最后可能在油体蛋白作用下形成油滴并储在于细胞中。ArA自油酸是开始经过多个去饱和酶及延长酶的作用而产生的。油酸是由硬脂酰 ACP去饱和酶作用而形成的,而棕榈油酸是在叶绿体的糖脂上被去饱和而产生的。三酰甘油最终被分解成自由脂肪酸和丙酮酸,而长链脂肪酸是以酰基 辅酶A的形式逐级降解的。
英文摘要:
      In order to understand the metabolic pathway of arachidonic acid and other lipids in Myrmecia incisa, the transcriptome pyrosequencing of this microalga was conducted by use of the sequencer Roche 454 GS FLX. Totally 393 722 reads (minimal size>29 bp) averaging 333 bp were generated from one consecutive pyrosequencing run. Cleaning of the raw sequences resulted in a total of 382 468 high quality reads with an average length of 322 nucleotides totalling 123 Mb. After clustering and assembly, these reads were assembled into 22 714 contigs and 25 621 singletons. The average length for contigs and singletons were 639 bp and 277 bp, respectively. By annotating the unisequences, the metabolic pathways of lipids were constructed. Fatty acid was de novo synthesized in chloroplasts, and free fatty acids were transported into cytosol where triacylglycerol was synthesized by endoplasmic reticulum. Oil bodies were formed possibly with the help of caleosins. Arachidonic acid was synthesized by desaturation for several times and elongation from oleic acid. Oleic acid was formed by stearoyl ACP desaturase, whereas palmitoic acid bound with glucolipid was generated by Δ7 desaturase. This research lays a foundation for systematic investigation into the manipulation of lipid metabolism and gene modification for higher production of ArA in M. incisa.
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