基于基因组和转录组的丹参酮生物合成相关基因SmCYP71AU66的筛选  

Identification of SmCYP71AU66 Related to Tanshinone Biosynthesis Based on Genome and Transcriptome of Salvia Miltiorrhiza

作　　者：于浩滢 高冉冉[2,3] 徐志超 闵伟红 Yu Haoying;Gao Ranran;Xu Zhichao;Min Weihong(College of Food Science and Engineering,Jilin Agricultural University,Changchun 130118,China;Engineering Research Center of Chinese Medicine Resource,Ministry of Education,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences/Peking Union Medical College,Beijing 100193,China;Key Lab of Chinese Medicine Resources Conservation,National Administration of Traditional Chinese Medicine of PeopleJ s Republic of China,Beijing 100193,China)

机构地区：[1]吉林农业大学食品科学与工程学院,长春130118 [2]中国医学科学院/北京协和医学院药用植物研究所中药资源教育部工程研究中心,北京100193 [3]国家中医药管理局中药资源保护重点研究室,北京100193

出　　处：《世界科学技术-中医药现代化》2019年第5期859-866,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金青年基金项目(31700264)：丹参酮生物合成关键酶短链脱氢酶编码基因的克隆和功能鉴定，负责人：徐志超

摘　　要：目的:以丹参基因组及全长转录组数据库为基础,对丹参酮生物合成关键酶基因进行预测,进一步推进丹参酮生物合成途径的解析。方法:结合丹参中CYP450s的系统发育鉴定丹参CYP71clan成员,采用生物信息技术分析其差异表达、基因结构和保守基序等;克隆候选基因并进行理化特性和蛋白结构等分析。结果:从丹参基因组注释到161个CYP71clan成员,分为16个家族,其中CYP71家族成员有64个,数量最多且结构域保守。转录组表达分析显示CYP71clan中36%的基因高表达(FPKM> 10)。克隆到基因SmCYP71AU66全长1503bp,编码500个氨基酸,在丹参根的周皮部位显著高表达,符合丹参酮在丹参体内的分布规律,预测该基因是丹参酮生物合成途径中的关键酶基因。结论:本研究为丹参酮合成相关基因的挖掘以及生物合成途径的解析提供参考。Objective: To predict the key enzyme gene involved in the biosynthesis of tanshinone based on the genome and full-length transcriptome database of Salvia miltiorrhiza, so as to further promote the pathway analysis in biosynthesis of tanshinone. Methods: The CYP71 clan members were identified via phylogenetic analysis of CYP450 s in Salvia miltiorrhiza, and the differential expression, gene structure, conserved motifs, etc were studied in silico. Candidate gene was cloned and analyzed its physicochemical properties and protein structure. Results: In total, 161 CYP71 clan members annotated in the genome of Salvia miltiorrhiza were classified into 16 families. Among them, 64 CYP450 s members belonged to CYP71 family, whose number was the largest and domain was conserved. Transcriptome expression analysis revealed that 36% of CYP71 clan genes were highly expressed with FPKM > 10. The SmCYP71 AU66 gene was cloned with the full-length of 1503 base pairs(bp), encoding 500 amino acids. SmCYP71 AU66 showed significantly high expression in the periderm of root, in accordance with the distribution of tanshinone, suggesting the key enzyme gene in tanshinone biosynthesis. Conclusion: This study provides the foundation for gene discovery and pathway analysis related to tanshinone biosynthesis.

关 键 词：丹参酮 生物合成 CYP71clan SmCYP71AU66 

分 类 号：R917[医药卫生—药物分析学]