The transcriptome pyrosequencing and gene function annotation of the green microalga Myrmecia incisa
The transcriptome pyrosequencing and gene function annotation of the green microalga Myrmecia incisa
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发表时间：
2012
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通讯作者：
Zhou Zhi-gang
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作者：
Zhou Zhi-gang
为了了解桃金娘体内花生四烯酸等脂类物质的代谢途径，采用Roche 454 GSFLX测序仪对该微卫星进行转录组焦磷酸测序，共获得393722个片段（最小大小29 bp）从一个连续的焦磷酸测序运行中产生了平均333 bp的序列。结果表明，该序列的平均长度为322个核苷酸，总长度为123 Mb，经聚类和拼接后，得到22 714个重叠群和25 621个单体序列，重叠群和单体序列的平均长度分别为639 bp和277 bp。油体的形成可能与钙蛋白有关;花生四烯酸由油酸经多次去饱和和延伸合成;油酸由硬脂酰-ACP去饱和酶合成，本研究为系统研究切割枝霉脂代谢调控和ArA高产基因修饰奠定了基础。
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 size29 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.