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苦参查尔酮异构酶的基因克隆与表达分析*(PDF)

《云南中医学院学报》[ISSN:1000-2723/CN:53-1048/R]

期数:
2019年03期
页码:
22-27
栏目:
实验研究
出版日期:
2020-04-20

文章信息/Info

Title:
Cloning and Expression Analysis of Chalcone Isomerase from Sophora flavescens
文章编号:
1000-2723(2019)03-0022-06
作者:
易雨琪韩荣春俞年军顾 晓苏 蝶童小慧许 菲王丽萍
(安徽中医药大学,安徽省中医药科学院中药资源保护与开发研究所,安徽 合肥 230012)
Author(s):
YI Yuqi HAN Rongchun YU Nianjun GU Xiao SU Die TONG Xiaohui XU Fei WANG Liping
(Anhui University of Chinese Medicine & Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine, Hefei 230012, China)
关键词:
苦参查尔酮异构酶基因克隆半定量PCR
Keywords:
Sophora flavescens chalcone isomerase gene cloning semi-quantitative PCR
分类号:
Q943.2;S567.239
DOI:
10.19288/j.cnki.issn.1000-2723.2019.03.004
文献标识码:
A
摘要:
目的为明晰苦参查尔酮异构酶(sfCHI)的结构基因信息及其在苦参黄酮类化合物生源路径中的表达调控机理。方法 本实验以苦参为研究对象,基于其转录组学数据,克隆sfCHI基因并针对该cDNA序列展开生物信息学分析,采用半定量PCR方法检测安徽产苦参不同组织中sfCHI基因表达情况。结果 结果显示苦参sfCHI基因的cDNA序列长度为1 265 bp,含有630 bp的开放式阅读框(ORF),编码209个氨基酸;序列分析表明sfCHI与狭叶羽扇豆查尔酮异构酶基因相似性最高,达89.95%;进化树分析该基因与大豆、野大豆等植物的亲缘关系较近;半定量PCR结果显示sfCHI基因在苦参根中表达量最低,在叶、叶柄与茎中的表达未见明显差异。结论 本研究通过对苦参查尔酮异构酶的基因克隆与不同组织间的表达分析,初步明确了该基因的序列信息与理化特征,这对进一步研究苦参黄酮类物质生物合成机制将起到积极的推动作用。
Abstract:
Objective In order to clarify the structural gene information of Sophora flavescens chalcone isomerase (sfCHI) and its expression pattern among distinct organs. Methods This experiment took advantage of S. flavescens deep transcriptomic data to clone sfCHI gene and conducted bioinformatics analysis on the cDNA sequence. Semi-quantitative PCR was used to detect the expression of sfCHI gene in different organs of S. flavescens. Results The results showed that the cDNA sequence of S. flavescens CHI gene was 1 265 bp in length and contained a 630 bp open reading frame (ORF) encoding 209 amino acids. Sequence analysis showed that the similarity between sfCHI and Lupinus angustifolius CHI gene was the highest, up to 89.95%. Phylogenetic tree resulted from 16 selected CHI genes showed that sfCHI and other related genes from Leguminosae were in the same group. Semi-quantitative PCR results showed that sfCHI gene expression in the root of S. flavescens is the lowest, while expression in leaves, petioles and stems did not show obvious difference. Conclusion In this study, the sequence information and physicochemical characteristics of the gene were firstly clarified by gene cloning of the chalcone isomerase gene and the expression analysis of different tissues, which will promote the study of biosynthetic mechanism of flavonoids in S. flavescens.

参考文献/References

[1] 顾观光辑.神农本草经[M]. 杨鹏举校注. 北京:学苑出版社,2007:143.
[2] 国家药典委员会. 中华人民共和国药典[S]. 北京:中国医药科技出版社,2015:202-203.
[3] HE X, FANG J, HUANG L, et al. Sophora flavescens Ait.:Traditional usage, phytochemistry and pharmacology of an important traditional Chinese medicine[J]. J Ethnopharmacol, 2015,172:10-29.
[4] 张翅,马悦,高慧敏,等. 苦参化学成分研究进展[J]. 中国实验方剂学杂志,2014,20(4):205-214.
[5] KUROYANAGI M, ARAKAWA T, HIRAYAMA Y, et al. Antibacterial and antiandrogen flavonoids from Sophora flavescens[J]. J Nat Prod,1999,62(12):1595-1599.
[6] 张钟媛. 苦参的化学成分和药理作用研究进展[J]. 云南中医中药杂志,2015,36(6):104-106.
[7] YIM D, KIM M J, SHIN Y, et al. Inhibition of cytochrome P450 activities by Sophora flavescens extract and its prenylated flavonoids in human liver microsomes[J]. Evid Based Complement Alternat Med,2019,2019:2673769.
[8] HILLERNS P I, WINK M. Binding of flavonoids from Sophora flavescens to the rat uterine estrogen receptor[J]. Planta Med,2005,71,(11):1065-1068.
[9] DENG X, SHEN C, MENG Q. Screening of herbal components for attenuating amiodarone-induced hepatotoxicity on gel-entrapped rat hepatocytes[J]. Drug Chem Toxicol,2014,37(1):100-106.
[10] 周明,沈勇根,朱丽琴,等. 植物黄酮化合物生物合成、积累及调控的研究进展[J]. 食品研究与开发,2016,37(18):216-220.
[11] JEZ J M, BOWMAN M E, DIXON R A, et al. Structure and mechanism of the evolutionary unique plant enzyme chalcone isomerase[J]. Nat Struct Biol,2000,7(9):786-791.
[12] YUAN Y, MA X, SHI Y, et al. Isolation and expression analysis of six putative structural genes involved in anthocyanin biosynthesis in Tulipa fosteriana[J]. Scientia Horticulturae,2013,153:93-102.
[13] GENSHEIMER M, MUSHEGIAN A. Chalcone isomerase family and fold: no longer unique to plants[J]. Protein Sci, 2004,13(2):540-544.
[14] BAN Z, QIN H, MITCHELL A J, et al. Noncatalytic chalcone isomerase-fold proteins in Humulus Lupulus are auxiliary components in prenylated flavonoid biosynthesis[J]. Proc Natl Acad Sci U S A,2018,115(22):E5223-E5232.
[15] 蔡雯婷,孙娟,董文华,等. 草莓果实查尔酮异构酶基因克隆及表达分析[J]. 北京农学院学报,2019,34(4):40-44.
[16] 王蕊,邹庆军,郭巧生,等. 杭菊CHI基因的克隆及原核表达[J]. 中国中药杂志,2019,44(14):3051-3021.
[17] 王蕊,李彦慧,郑健. 紫丁香查尔酮异构酶基因SoCHI的克隆及表达分析[J]. 植物资源与环境学报,2018,27(3):11-17.
[18] HAN R, TAKAHASHI H, NAKAMURA M, et al. Transciptome analysis of nine tissues to discover genes involved in the biosynthesis of active ingredients in Sophora flavescents[J]. Biol Pharm Bull,2015,38(6):876-883.
[19] MAGRANE M, CONSORTIUM U. UniProt Knowledgebase:a hub of integrated protein data[J]. Database(Oxford),2011,2011:bar009.
[20] KUMAR S, STECHER G, LI M, et al. MEGA X:Molecular Evolutionary Genetics Analysis across computing platforms[J]. Mol Biol and Evol,2018,35(6):1547-1549.
[21] MCGUFFIN L J, ADIYAMAN R, MAGHRABI A H, et al. IntFOLD:an integrated web resource for high performance protein structure and function prediction[J]. Nucl Acids Res,2019,47(W1):W408-W413.
[22] MCGUFFIN L J, SHUID A N, KEMPSTER R, et al. Accurate template-based modeling in CASP12 using the IntFOLD4-TS,ModFOLD6,and ReFOLD methods[J]. Proteins,2018,86(S1):335-344.

备注/Memo

备注/Memo:
收稿日期: 2019 - 04- 18
* 基金项目: 国家重点研发计划中医药现代化研究重点专项(2017YFC1701602);安徽省高校优秀青年人才支持计划项目(gxyq2017016);安徽省重点研究与开发计划项目(1804b06020356);安徽中医药大学自然重点项目(2017zrzd011,2019zrzd02)
第一作者简介: 易雨琪(1997-),女,在读硕士研究生,研究方向:药用植物次生代谢产物生物合成路径研究。
△通信作者: 韩荣春, E-mail: hanr@ahtcm.edu.cn
更新日期/Last Update: 2020-05-06