浙贝母肌动蛋白基因的克隆及生物信息学分析

发布时间：2018-03-04 12:13

本文选题：浙贝母　切入点：肌动蛋白(Actin)基因　出处：《中成药》2017年01期 　论文类型：期刊论文

【摘要】：目的克隆浙贝母Fritillaria thunbergii Miq.肌动蛋白(Actin)基因,并进行生物信息学分析。方法提取浙贝母根、茎、叶、花、鳞茎总RNA,设计合成简并引物。以叶片总RNA为模板,通过RT-PCR和Ta克隆技术克隆Actin基因保守区序列片段。以该基因为内参基因,对3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)基因进行组织特异性表达分析。结果经RT-PCR扩增和Ta克隆,得到1段基因序列(463 bp)。浙贝母Actin基因与岷江百合、郁金香、虎眼万年青、铁皮石斛、柿子、光皮桦、玉米相似度较高(84%~98%),其氨基酸序列与阿帝露茅膏菜、甘蓝型油菜、香草兰、岷江百合、麻疯树、枸杞、红海榄的同源性均在89%以上,并且其Actin蛋白与百脉根、岷江百合、郁金香亲缘关系较近。HMGR基因在该植物各部位表达有显著差异,依次为鳞茎花叶茎根。结论首次从浙贝母中克隆出Actin基因(命名为Ft Actin),可为其有效利用奠定基础。
[Abstract]:Objective to clone Fritillaria thunbergii Miq. actin (Actinin) gene and analyze it by bioinformatics. Methods Total RNAs of root, stem, leaf, flower and bulb of Fritillaria thunbergii were extracted, and degenerate primers were designed and synthesized. Total RNA of leaves was used as template. The conserved sequence of Actin gene was cloned by RT-PCR and Ta cloning techniques. The 3- hydroxy-3-methyl-glutaryl coA reductase (HMGR) gene was analyzed for tissue specific expression by RT-PCR amplification and Ta cloning. A fragment of Actin gene of Fritillaria thunbergii was obtained, and its amino acid sequence was similar to that of Lilium tinctorius, tulip, evergreen tiger eye, Dendrobium candidum, Persimmon, Betula lucifera and maize. The amino acid sequence was similar to that of Artemisia, Brassica napus, vanilla orchid, and its amino acid sequence was similar to that of Artilla, Brassica napus and Vanilla. The homology of Minjiang Lilium, Jatropha curcas, Lycium barbarum and Red Sea Elm were all above 89%, and there were significant differences in the expression of its Actin protein with the close relationship between Lilium officinalis, Lilium Minjiang and Tulip in all parts of the plant. Conclusion the Actin gene (named Ft Actinus) was cloned from Fritillaria thunbergii for the first time, which could lay a foundation for its effective utilization.
【作者单位】：浙江万里学院生物技术研究所;浙江省中药研究所;
【基金】：浙江省中药材育种专项(2016C02058) 宁波市公益类重大专项(2015C110007) 浙江万里学院重中之重学科学生创新项目(CX2015002)
【分类号】：S567.231

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