濒危药用植物茅苍术法呢基焦磷酸合酶基因克隆及其表达分析

发布时间：2018-09-13 08:51

【摘要】：目的克隆茅苍术Atractylodes lancea倍半萜类成分生物合成关键酶法呢基焦磷酸合酶(FPPS)基因全长并分析其表达规律。方法以茅苍术总RNA为模板,运用同源克隆法和RACE技术克隆茅苍术FPPS基因(Al FPPS)的c DNA全长,并进行生物信息学分析;用q RT-PCR和GC-MS分析不同生长阶段FPPS在茅苍术根茎中的表达及倍半萜类成分量变化,并分析二者的相关性。结果获得Al FPPS基因全长c DNA为1 320 bp(Gen Bank accession no.KX443242),含一个长1 029 bp的开放阅读框,编码342个氨基酸,包含5个保守功能域,其中2个富含天冬氨酸(DXXDD)。q RT-PCR及GC-MS结果显示不同时期茅苍术Al FPPS基因表达量与倍半萜类成分量呈显著正相关。结论首次克隆获得Al FPPS基因的全长c DNA,初步证明Al FPPS基因是茅苍术倍半萜类成分生物合成途径中的重要调控位点,为茅苍术倍半萜类成分生物合成途径阐明与生物工程应用提供科学依据。
[Abstract]:Objective to clone the full length of the (FPPS) gene of Atractylodes lancea sesquiterpene biosynthesis and analyze its expression pattern. Methods using total RNA of Atractylodes macrocephala as template, the length of c DNA of FPPS gene (Al FPPS) was cloned by homologous cloning method and RACE technique, and bioinformatics analysis was carried out. Q RT-PCR and GC-MS were used to analyze the expression of FPPS in Rhizome of Atractylodes macrocephala and the changes of sesquiterpene components in the rhizome of Atractylodes macrocephala, and the correlation between them was analyzed. Results the full-length c DNA of Al FPPS gene was 1 320 bp (Gen Bank accession no.KX443242), containing an open reading frame of 1 029 bp, encoding 342 amino acids and containing 5 conserved functional domains. Two of them were rich in aspartic acid (DXXDD). Q RT-PCR and GC-MS. The results showed that there was a significant positive correlation between Al FPPS gene expression and sesquiterpenoid components in different periods. Conclusion the full length c DNA, of Al FPPS gene was cloned for the first time. It is proved that Al FPPS gene is an important regulatory site in the biosynthesis pathway of sesquiterpenoid components of Atractylodes macrocephala. It provides a scientific basis for elucidating the biosynthesis pathway of sesquiterpenoids and the application of bioengineering in Rhizoma Atractylodes.
【作者单位】：南京中医药大学药学院;
【基金】：国家自然科学基金资助项目(81573520) 江苏省中药优势学科II期建设(ysxk-2014) “六大人才高峰”高层次人才项目(2012-YY-009) 江苏省“333高层次人才培养工程” 江苏省中药资源产业化过程协同创新中心资助项目
【分类号】：S567.239

【相似文献】