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利用Illumina的Genome Analyzer IIx对银杏(Ginkgo Biloba)细胞转录组进行高通量测序,挖掘银杏内酯和紫杉醇生物合成基因,特别是新的羟基化酶基因,为今后最终完善红豆杉细胞紫杉醇生物合成途径中未知的羟基化步骤作准备。通过测序,获得了银杏细胞69 286个contig,56 387个scaffold,32 032个unigene。Unigene平均长度636bp。另外从gap分布、GC含量、基因组coverage等方面对unigene进行评估,数据显示测序质量好,可信度高。通过分析unigene的表达和功能注释等信息,发现66个属于CYP450基因家族,726个参与次生代谢物合成,其中59个与萜类合成有关,17个与二萜类合成相关。利用生物信息学方法从Michigan State University银杏成熟叶、侧根、成熟果实、无菌苗以及次生茎的转录组数据中找到了与银杏细胞CYP450高度同源的紫杉烷羟基化酶候选基因15个,为后续研究奠定了基础。
High-throughput sequencing of Ginkgo Biloba cell transcriptome using Genome Analyzer IIx from Illumina revealed that Ginkgo biloba lactone and paclitaxel biosynthesis genes, especially the new hydroxylase gene, could be successfully excised for future paclitaxel biosynthesis Route unknown hydroxylation step in preparation. By sequencing, 69 286 contigs, 56 387 scaffolds and 32 032 unigene were obtained. Unigene average length of 636bp. In addition, unigene was evaluated in terms of gap distribution, GC content and genome coverage. The data showed good sequencing quality and high credibility. By analyzing information such as unigene expression and functional annotation, we found that 66 belong to the CYP450 gene family and 726 are involved in the synthesis of secondary metabolites, of which 59 are related to terpenoid synthesis and 17 are related to diterpenoid synthesis. Bioinformatics methods were used to find fifteen candidate taxane hydroxylase genes that were highly homologous to CYP450 of Ginkgo biloba from the transcriptome data of mature leaves, lateral roots, mature fruits, sterile seedlings and secondary stems of Michigan State University. , Laid the foundation for follow-up research.