| 吕楠 安红强 裴薇 王万军△.铁皮石斛尿苷二磷酸葡萄糖焦磷酸化酶基因的克隆与分析[J].现代生物医学进展英文版,2017,17(7):1215-1219. |
| 铁皮石斛尿苷二磷酸葡萄糖焦磷酸化酶基因的克隆与分析 |
| Cloning and Expression Analysis of UGPase Gene in DendrobiumOfficinaleKimura et Migo |
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| DOI： |
| 中文关键词: 铁皮石斛多糖 尿苷二磷酸葡萄糖焦磷酸化酶 qRT-PCR 蛋白结构 |
| 英文关键词: Dendrobiumpolysaccharides DoUGPase1 qRT-PCR Protein structure |
| 基金项目:国家自然科学基金项目(31371232) |
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| 中文摘要: |
| 目的：研究尿苷二磷酸葡萄糖焦磷酸化酶（UGPase）在铁皮石斛不同组织中的表达情况，探讨其与石斛多糖累积的关系。方
法：根据铁皮石斛原球茎转录组数据中的DoUGPase1 序列设计引物，并利用RACE 技术克隆该基因；用生信分析软件预测其蛋
白结构，并进行系统发育分析；利用实时荧光定量PCR (qRT-PCR)技术研究该基因在铁皮石斛不同年份，不同组织中的表达。结
果：通过RACE技术得到1个3054 bp 基因，命名为DoUGPase1，其ORF长1530 bp，编码510 个氨基酸残基，与油棕(Elaeis
guineensis)和波斯枣(Phoenix dactylifera)有一定的同源性，且DoUGPase1 在3 年生的植株中相对表达量整体较高(P<0.05)。结论：
我们预测在铁皮石斛的生长发育过程中DoUGPase1 基因与多糖累积的活跃程度成正比例。 |
| 英文摘要: |
| Objective:To observe the expression of UGPase in Dendrobium, and investigate its relationship with Dendrobium
polysaccharide accumulation.Methods:The primers were designed according to the transcriptomic UGPase1 unigene sequence of protocormin
D. officinale, then DoUGPase1 transcription factors were cloned by RACE techniques. Through bioinformatics analysis and phylogenesis
analysis, the protein structure was predicted, and its affinity was obstained. The relative expression of D.officinale in different
years and different tissues was analyzed through Roche Light circle 96 real-time fluorescent quantitative PCR (RT-qPCR) experiment.Results:By RACE technique, a new 1530 bp long gene named DoUGPase1 was obstained. Its ORF (Open reading frame) was 1530 bp
encoding 510 amino acids. The DoUGPase1 had the relatives with Elaeis guineensis and Phoenix dactylifera. The DoUGPase1 expression
in 3 years plant was the biggest（P<0.05）Conclusion:The expression level of DoUGPase1gene might be a positive proportion with
active degree of dendrobiumpolysaccharide accumulation in the growth and development of dendrobiumofficinale. |
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