紫金龙异紫堇定生物合成相关6-OMT基因克隆与功能表征

doi:10.13560/j.cnki.biotech.bull.1985.2024-0544

生物技术通报 ›› 2025, Vol. 41 ›› Issue (2): 309-320.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0544

• 研究报告 • 上一篇

紫金龙异紫堇定生物合成相关6-OMT基因克隆与功能表征

李明¹^,²^,³(), 刘祥宇¹^,²^,³, 王益娜¹^,²^,³, 和四梅¹^,²^,³^,⁴, 沙本才²^,³^,⁴()

^1.云南农业大学农学与生物技术学院，昆明 650201
^2.云南农业大学西南中药材种质创新与利用国家地方联合工程研究中心，昆明 650201
^3.云南农业大学云南省药用植物生物学重点实验室，昆明 650201
^4.云南特色植物提取实验室，昆明 650106

收稿日期:2024-06-07 出版日期:2025-02-26 发布日期:2025-02-28
通讯作者: 沙本才，男，副教授，研究方向：作物栽培与耕作；E-mail: ynshbc@aliyun.com
作者简介:李明，男，硕士研究生，研究方向：药用植物生物合成；E-mail: 3049730657@qq.com
基金资助:
国家自然科学基金项目(82160727);云南特色植物提取实验室自主研究项目基金(2022YKZY001)

Cloning and Functional Characterization of 6-OMT Gene Related to Isocorydine Biosynthesis in Dactylicapnos scandens

LI Ming¹^,²^,³(), LIU Xiang-yu¹^,²^,³, WANG Yi-na¹^,²^,³, HE Si-mei¹^,²^,³^,⁴, SHA Ben-cai²^,³^,⁴()

^1.College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201
^2.National Local Joint Engineering Research Center for Germplasm Innovation and Utilization of Chinese Medicinal Materials in Southwest, Yunnan Agricultural University, Kunming 650201
^3.Yunnan Key Laboratory of Medicinal Plant Biology, Yunnan Agricultural University, Kunming 650201
^4.Yunnan Characteristic Plant Extraction Laboratory, Kunming 650106

Received:2024-06-07 Published:2025-02-26 Online:2025-02-28

摘要/Abstract

摘要：

目的（S）-去甲乌药碱O-甲基转移酶（6-OMT）是异紫堇定生物合成的关键限速酶，通过克隆与体外酶活验证紫金龙6-OMT基因功能，为解析紫金龙异紫堇定生物合成途径奠定基础。方法从紫金龙转录组数据中挖掘DsOMT基因，通过PCR扩增获得全长cDNA序列，并通过生物信息学分析DsOMT蛋白结构；分析DsOMT基因在不同组织中的表达水平；构建pET-28a-DsOMT原核表达载体，将其转入大肠杆菌BL21（DE3）进行诱导表达，纯化蛋白后进行体外酶反应，以表征其功能。结果从转录组数据中挖掘到4个DsOMT候选基因，分别命名为DsOMT07、DsOMT08、DsOMT010、DsOMT012，并成功扩增得到全长cDNA序列；4个DsOMT蛋白都不存在跨膜结构域和信号肽，属于膜外蛋白。系统发育显示4个DsOMT与6-OMT亚家族亲缘关系较近。对4个DsOMT的氨基酸序列进行分析，显示可能具有上游途径（S）-去甲乌药碱6-OH位点的催化活性。表达谱分析发现4个DsOMT基因在根中高表达。SDS-PACE结果表明DsOMT蛋白可溶性高，并在大肠杆菌中高效表达，体外酶反应后发现DsOMT010能够催化（S）-去甲乌药碱的C6的O-甲基化形成（S）-衡州乌药碱。结论成功克隆了4个DsOMT基因，属于膜外蛋白，与6-OMT亚家族亲缘关系较近，4个DsOMT基因在根中高表达；同时在大肠杆菌中实现了DsOMT的异源表达，并纯化蛋白进行了体外酶功能表征，鉴定到1个（S）-去甲乌药碱C6位O-甲基转移酶DsOMT010。

关键词: 紫金龙, O-甲基转移酶, 生物信息学分析, 原核表达, 功能表征

Abstract:

Objective (S)-norcoclaurine O-methyltransferase (6-OMT) is the key rate-limiting enzyme in the biosynthesis of isocorydine. This study is aimed to verify the function of the 6-OMT gene from Dactylicapnos scandens through cloning and in vitro enzyme activity assays, thereby laying the foundation for elucidating the isocorydine biosynthetic pathway in D. scandens. Method The DsOMT gene was mined from the transcriptome data of D. scandens. The full-length cDNA sequence was obtained via PCR amplification, and the protein structure of DsOMT was analyzed using bioinformatics. The expressions of the DsOMT gene in different tissues were examined. A pET-28a-DsOMT prokaryotic expression vector was constructed and transferred into Escherichia coli BL21(DE3) for induced expression. The protein was then purified and subjected to in vitro enzyme assays to characterize its function. Result Four DsOMT candidate genes, named DsOMT07, DsOMT08, DsOMT010, and DsOMT012, were identified from the transcriptome data, and their full-length cDNA sequences were successfully amplified. All four DsOMT proteins lacked transmembrane domains and signal peptides, classifying them as extramembrane proteins. Phylogenetic analysis indicated that these genes were closely related to the 6-OMT subfamily. Amino acid sequence analysis suggested potential catalytic activity at the 6-OH site of the upstream pathway for (S)-norcoclaurine. Expression profiling revealed that these four DsOMT genes were highly expressed in the roots. SDS-PAGE results showed that DsOMT proteins were highly soluble and efficiently expressed in Escherichia coli. In vitro enzyme assays demonstrated that DsOMT010 catalyzed the O-methylation of the C6 position of (S)-norcoclaurine, forming (S)-coclaurine. Conclusion Four DsOMT genes are successfully cloned and identified as extramembrane proteins closely related to the 6-OMT subfamily. These genes show high expressions in the roots. Additionally, heterologous expression of DsOMT is achieved in E. coli, and the purified proteins are characterized for their in vitro enzyme functions. Among them, DsOMT010 is identified as a (S)-norcoclaurine C-6 O-methyltransferase.

Key words: Dactylicapnos scandens, O-methyltransferase, bioinformatics analysis, prokaryotic expression, functional characterization

李明, 刘祥宇, 王益娜, 和四梅, 沙本才. 紫金龙异紫堇定生物合成相关6-OMT基因克隆与功能表征[J]. 生物技术通报, 2025, 41(2): 309-320.

LI Ming, LIU Xiang-yu, WANG Yi-na, HE Si-mei, SHA Ben-cai. Cloning and Functional Characterization of 6-OMT Gene Related to Isocorydine Biosynthesis in Dactylicapnos scandens[J]. Biotechnology Bulletin, 2025, 41(2): 309-320.

图/表 13

图1 紫金龙各组织样及化合物结构A-C分别为采集的紫金龙根、茎、叶组织；D和E分别为异紫堇定碱和（S）-去甲乌药碱的结构及位置编号

Fig. 1 Structure of tissue samples and compounds of D. scandensA-C refers to the roots, stems and leaves of D. scandens, respectively. D and E are the structure and position numbers of isovioladinine and (S)-norcoclaurine, respectively

表1 生物信息学分析网站链接

Table 1 Website links on bioinformatics analysis

项目列表 Analysis item	网址 Website
NCBI同源基因比对分析	https：//blast.ncbi.nlm.nih.gov/Blast.cgi
蛋白理化性质分析	https：//web.expasy.org/protparam/
蛋白信号肽预测	https：//services.healthtech.dtu.dk/services/SignalP-6.0/
蛋白跨膜结构域预测	https：//services.healthtech.dtu.dk/services/TMHMM-2.0/
蛋白二级结构预测	https：//npsa-prabi.ibcp.fr/cgi-bin/npsa
蛋白三级结构预测	https：//swissmodel.expasy.org/
蛋白保守基序分析	https：//meme-suite.org/meme/
蛋白保守结构域分析	https：//www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi
蛋白亚细胞定位	http：//www.csbio.sjtu.edu.cn/bioinf/Cell-PLoc-2/
多序列比对	https：//espript.ibcp.fr/ESPript/cgi-bin/ESPript.cgi

表2 PCR引物序列

Table 2 PCR primer sequences

基因 Gene	正向引物序列 Forward primer sequence （5′-3′）	反向引物序列 Reverse primer sequence （5′-3′）
DsOMT08	cagcaaatgggtcgcggatccATGGAAGTGAAGAAGAGTGAAC	gacggagctcgaattcggatccTTAATATGGATAAACCTCAATAACAG
DsOMT07	cagcaaatgggtcgcggatccATGGAGATGATCATCAATGAAG	acggagctcgaattcggatccCTAAGGATATGCAACAATAACTG
DsOMT010	cagcaaatgggtcgcggatccATGGGTGTCAGTGATATAGCTG	acggagctcgaattcggatccCTATGGGAAGGCTTCAATGACAG
DsOMT012	cagcaaatgggtcgcggatccATGGGTGCCAATGAGATAGCAG	acggagctcgaattcggatccCTATGGGAAGGCTTCAATGACAG
PCR-pET-28a	taatacgactcactataggggaatt	aaacgggtcttgaggggttttttg

图2 紫金龙候选基因热图、与其他植物的甲基转移酶基因的系统进化树及保守基序分析A：MT家族进化树分析；B：候选基因在紫金龙不同组织中的表达量热图；C：候选基因与其他物种的OMT基因的进化树（前）和保守基序（后）分析

Fig. 2 Candidate gene heat map of D. scandens, phylogenetic tree and conserved motif analysis of methyltransferase genes in other plantsA: Analysis of MT family phylogenetic tree. B: Expression heat map of candidate genes in different tissues of D. scandens. C: Phylogenetic tree (pre) and conserved motif (post) analysis of candidate genes and OMT genes of other species

图3 四个筛选的DsOMT基因与其他物种OMT基因的多序列比对

Fig. 3 Multiple sequence alignment of four screened DsOMT genes with OMT genes of other species

表3 DsOMT蛋白的理化性质预测统计

Table 3 Prediction and statistics of physicochemical properties of DsOMT protein

基因 Gene	氨基酸数目 Amino acid number	分子量 Molecular weight/Da	理论等电点 Theoretical pI	不稳定系数 Instability index	脂肪系数 Fat coefficient	亲水系数 Hydrophilicity coefficient	亚细胞定位 Subcellular localization
DsOMT07	356	39 577.72	4.95	28.65	105.96	0.024	叶绿体
DsOMT08	353	39 349.51	5.61	33.31	93.34	-0.150	叶绿体
DsOMT010	349	38 582.51	5.23	36.41	102.23	0.034	叶绿体
DsOMT012	348	38 346.22	5.47	38.58	102.53	0.025	叶绿体

图4 四个筛选的DsOMT蛋白跨膜结构域（上）及信号肽预测（下）A-D为DsOMT07、DsOMT08、DsOMT010和DsOMT012蛋白的跨膜结构域（上）及信号肽（下）预测

Fig. 4 Transmembrane structural domains of four screened DsOMT proteins (top) and signaling peptide prediction (bottom)A-D are the predicted transmembrane structural domains (top) and signaling peptides (bottom) of DsOMT07, DsOMT08, DsOMT010, and DsOMT012 proteins

图5 四个DsOMT蛋白的二级结构预测A-D依次是DsOMT07、DsOMT08、DsOMT010和DsOMT012蛋白二级结构预测；E：四个DsOMT蛋白的二级结构预测统计

Fig. 5 Secondary structure prediction of four DsOMT proteinsA-D are the predicted secondary structures of DsOMT07, DsOMT08, DsOMT010 and DsOMT012 proteins in order. E: Secondary structure prediction statistics of four DsOMT proteins

图6 四个DsOMT蛋白的三级结构预测A-D依次为DsOMT07、DsOMT08、DsOMT010和DsOMT012蛋白三级结构预测；蓝色：螺旋；绿色：延伸链；银白色：无规则卷曲

Fig. 6 Prediction of the tertiary structure of four DsOMT proteinsA-D predicted tertiary structure of DsOMT07, DsOMT08, DsOMT010 and DsOMT012 proteins in order; blue: spiral; green: extension chain; silver-white: irregular curl

图7 紫金龙总RNA提取及DsOMT基因的克隆A：紫金龙各组织总RNA电泳胶图，泳道1-3分别为紫金龙根、茎、叶组织总RNA电泳胶图；B：克隆4个候选的DsOMT基因电泳胶图，泳道4-7分别为DsOMT07、DsOMT08、DsOMT010、DsOMT012基因克隆电泳胶图；泳道M：DNA marker 5 000电泳条带

Fig. 7 Extraction of total RNA from D. scandens and cloning of DsOMT geneA: Total RNA electrophoresis gel diagram of each tissue of D. scandens, lane 1-3 are total RNA electrophoresis gel diagram of root, stem and leaf tissues of D. scandens, respectively. B: Clone 4 candidate DsOMT gene electrophoresis gels, lane 4-7are cloned for DsOMT07, DsOMT08, DsOMT010 and DsOMT012 genes, respectively. Lane M: DNA marker 5 000 electrophoresis band

图8 pET-28a载体单酶切及DsOMT重组pET-28a验证A：单酶切的pET-28a线性载体电泳胶图，泳道1为未酶切的pET-28a载体，泳道2和3为已酶切的pET-28a载体；B：菌液检测阳性的pET-28a-DsOMT（+）载体电泳胶图，泳道4-5、6-7、8-9和10-11分别为pET-28a-DsOMT 07（+）、pET-28a-DsOMT 08（+）、pET-28a-DsOMT 010（+）和pET-28a-DsOMT 012（+）菌液验证电泳胶图；泳道M为DNA marker 5 000电泳条带

Fig. 8 Single enzyme digestion of pET-28a vector and verification of DsOMT recombinant pET-28aA: Gel diagram of single-digested pET-28a linear vector, lane 1 is the undigested pET-28a vector, and lane 2 and 3 are the digested pET-28a vector. B: Electrophoresis gel diagram of pET-28a-DsOMT(+) vector with positive bacterial water test, and pET-28a-DsOMT 07(+), pET-28a-DsOMT 08(+), pET-28a-DsOMT 010(+) and pET-28a-DsOMT 012(+) in lane 4-5, 6-7, 8-9 and 10-11, respectively. Lane M is the DNA marker 5 000 electrophoresis band

图9 DsOMT蛋白纯化A-D分别为DsOMT07、DsOMT08、DsOMT010、DsOMT012蛋白250 mmol/L咪唑纯化的电泳胶图；A-D中泳道1-3为重复的蛋白样品条带，图A、C和D中的泳道M为蛋白marker 150，图B中的泳道M为蛋白marker 310

Fig. 9 Purification of DsOMT proteinA-D are electropherograms of 250 mmol/L imidazole purification of DsOMT07, DsOMT08, DsOMT010 and DsOMT012 proteins, respectively. Lane 1-3 in A-D are duplicate protein sample bands, lane M in panel A, C, and D is protein marker 150, and lane M in panel B is protein marker 310

图10 蛋白体外酶反应及LC-MS检测A：DsOMT010体外酶反应的催化步骤；B：LC-MS检测分析的TIC图；C：LC-MS检测的（S）-衡州乌药碱标准品EIC质谱图；D：LC-MS检测DsOMT010+（S）-去甲乌药碱的产物EIC质谱图

Fig. 10 Protein enzymatic reaction in vitro and LC-MS detectionA: Catalytic steps of DsOMT010 in vitro enzymatic reaction. B: TIC diagram of LC-MS detection and analysis. C: LC-MS detection of (S)-coclaurine standard EIC mass spectrometry. D: LC-MS detection of DsOMT010+(S)-norcoclaurine product EIC mass spectrometry

参考文献 44

1	中国科学院中国植物志编辑委员会. 中国植物志-第32卷[M]. 北京: 科学出版社, 1999.
	Editorial Committee of Flora of China Chinese Academy of Sciences. Flora of China (Vol. 32) [M]. Beijing: Science Press, 1999.
2	罗建蓉, 钱金栿, 周浓. 紫金龙脂溶性化学成分的研究 [J]. 西北药学杂志, 2010, 25(1): 19-20.
	Luo JR, Qian JF, Zhou N. Analysis of the hydrophobic components in Dactylicapnos scandens by GC-MS [J]. Northwest Pharm J, 2010, 25(1): 19-20.
3	张伟, 丁宏莉, 张斌. RP-HPLC法同时测定紫金龙中2种生物碱的含量 [J]. 云南中医学院学报, 2009, 32(6): 18-19, 47.
	Zhang W, Ding HL, Zhang B. Simultaneous RP-HPLC determination of two alkaloids in root of Dactylicapnos scandens [J]. J Yunnan Univ Tradit Chin Med, 2009, 32(6): 18-19, 47.
4	吴勐, 王银叶, 艾铁民. 紫金龙总生物碱的镇痛作用及其机制初探 [J]. 中草药, 2003, 34(11): 1022-1025.
	Wu M, Wang YY, Ai TM. Studies of analgesic effect and mechanism of total alkaloid extracted from Dactylicapnos scandens [J]. Chin Tradit Herb Drugs, 2003, 34(11): 1022-1025.
5	Guo CC, Jiang Y, Li L, et al. Application of a liquid chromatography-tandem mass spectrometry method to the pharmacokinetics, tissue distribution and excretion studies of Dactylicapnos scandens in rats [J]. J Pharm Biomed Anal, 2013, 74: 92-100.
6	Wang X, Dong HJ, Yang B, et al. Preparative isolation of alkaloids from Dactylicapnos scandens using pH-zone-refining counter-current chromatography by changing the length of the separation column [J]. J Chromatogr B, 2011, 879(31): 3767-3770.
7	王富华, 胡晓, 陈海林, 等. 紫金龙的生物碱类成分 [J]. 中国中药杂志, 2009, 34(16): 2057-2059.
	Wang FH, Hu X, Chen HL, et al. Alkaloids from Dactylicapnos scandens hutch [J]. China J Chin Mater Med, 2009, 34(16): 2057-2059.
8	张文鑫, 韩丽妹, 周毅生, 等. RP-HPLC同时测定紫金龙中4种异喹啉生物碱的含量 [J]. 中国药学杂志, 2009, 44(3): 233-236.
	Zhang WX, Han LM, Zhou YS, et al. Simultaneous determination of four isoquinoline alkaloids in root of Dactylicapnos scandens by RP-HPLC [J]. Chin Pharm J, 2009, 44(3): 233-236.
9	刘金凤, 黄鹏, 卿志星, 等. 苄基异喹啉类生物碱生物合成与代谢工程研究进展 [J]. 基因组学与应用生物学, 2016, 35(8): 2194-2200.
	Liu JF, Huang P, Qing ZX, et al. Advances in the biosynthesis and metabolic engineering research of benzylisoquinoline alkaloids [J]. Genom Appl Biol, 2016, 35(8): 2194-2200.
10	Jiang B, Gao L, Wang HJ, et al. Characterization and heterologous reconstitution of Taxus biosynthetic enzymes leading to baccatin III [J]. Science, 2024, 383(6683): 622-629.
11	Zhou C, Chen TJ, Gu AD, et al. Combining protein and metabolic engineering to achieve green biosynthesis of 12β-O-Glc-PPD in Saccharomyces cerevisiae [J]. Green Chem, 2023, 25(4): 1356-1367.
12	Paddon CJ, Westfall PJ, Pitera DJ, et al. High-level semi-synthetic production of the potent antimalarial artemisinin [J]. Nature, 2013, 496(7446): 528-532.
13	辛爱一, 柳军玺, 邸多隆. 阿朴菲类生物碱研究进展 [J]. 中草药, 2018, 49(3): 712-724.
	Xin AY, Liu JX, Di DL. Research progress on aporphine alkaloids [J]. Chin Tradit Herb Drugs, 2018, 49(3): 712-724.
14	Han JN, Li SJ. De novo biosynthesis of berberine and halogenated benzylisoquinoline alkaloids in Saccharomyces cerevisiae [J]. Commun Chem, 2023, 6(1): 27.
15	Ikezawa N, Iwasa K, Sato F. Molecular cloning and characterization of CYP80G2, a cytochrome P450 that catalyzes an intramolecular C-C phenol coupling of (S)-reticuline in magnoflorine biosynthesis, from cultured Coptis japonica cells [J]. J Biol Chem, 2008, 283(14): 8810-8821.
16	Leng L, Xu ZC, Hong BX, et al. Cepharanthine analogs mining and genomes of Stephania accelerate anti-coronavirus drug discovery [J]. Nat Commun, 2024, 15(1): 1537.
17	Zhang LB, Zhang SQ, Liao LJ, et al. Discovery, structure, and mechanism of the (R, S)-norcoclaurine synthase for the chiral synthesis of benzylisoquinoline alkaloids [J]. ACS Catal, 2023, 13(22): 15164-15174.
18	Farrow SC, Hagel JM, Beaudoin GAW, et al. Stereochemical inversion of (S)-reticuline by a cytochrome P450 fusion in opium poppy [J]. Nat Chem Biol, 2015, 11(9): 728-732.
19	Yang Y, Yuan JS, Ross J, et al. An Arabidopsis thaliana methyltransferase capable of methylating farnesoic acid [J]. Arch Biochem Biophys, 2006, 448(1/2): 123-132.
20	王志刚, 吴建新. DNA甲基转移酶分类、功能及其研究进展 [J]. 遗传, 2009, 31(9): 903-912.
	Wang ZG, Wu JX. DNA methyltransferases: classification, functions and research progress [J]. Hereditas, 2009, 31(9): 903-912.
21	Schmidberger JW, James AB, Edwards R, et al. Halomethane biosynthesis: Structure of a SAM-dependent halide methyltransferase from Arabidopsis thaliana [J]. Angew Chem Int Edit. 2010, 49(21): 3646-3648.
22	Lin Z, Hu ZW, Zhou LJ, et al. A large conserved family of small-molecule carboxyl methyltransferases identified from microorganisms [J]. Proc Natl Acad Sci U S A, 2023, 120(20): e2301389120.
23	Männistö PT, Kaakkola S. Catechol-O-methyltransferase (COMT): Biochemistry, molecular biology, pharmacology, and clinical efficacy of the new selective COMT inhibitors [J]. Pharmacol Rev, 1999, 51(4): 593-628.
24	Stanevich V, Jiang L, Satyshur KA, et al. The structural basis for tight control of PP2A methylation and function by LCMT-1 [J]. Mol Cell, 2011, 41(3): 331-342.
25	Li BZ, Yuan YJ, Jia B. Prospects of synthetic biology [J]. Sci Sin Chim, 2014, 44(9): 1455-1460.
26	Dang TTT, Facchini PJ. Characterization of three O-methyltransferases involved in noscapine biosynthesis in opium poppy [J]. Plant Physiol, 2012, 159(2): 618-631.
27	Chang LM, Hagel JM, Facchini PJ. Isolation and characterization of O-methyltransferases involved in the biosynthesis of glaucine in Glaucium flavum [J]. Plant Physiol, 2015, 169(2): 1127-1140.
28	Bu JL, Zhang XH, Li QS, et al. Catalytic promiscuity of O-methyltransferases from Corydalis yanhusuo leading to the structural diversity of benzylisoquinoline alkaloids [J]. Hortic Res, 2022, 9: uhac152.
29	Menéndez-Perdomo IM, Facchini PJ. Isolation and characterization of two O-methyltransferases involved in benzylisoquinoline alkaloid biosynthesis in sacred lotus (Nelumbo nucifera) [J]. J Biol Chem, 2020, 295(6): 1598-1612.
30	Chang JJ, Guo YL, Yan JY, et al. The role of watermelon caffeic acid O-methyltransferase (ClCOMT1) in melatonin biosynthesis and abiotic stress tolerance [J]. Hortic Res, 2021, 8: 210.
31	Jin JQ, Qu FR, Huang HS, et al. Characterization of two O-methyltransferases involved in the biosynthesis of O-methylated catechins in tea plant [J]. Nat Commun, 2023, 14(1): 5075.
32	He SM, Song WL, Cong K, et al. Identification of candidate genes involved in isoquinoline alkaloids biosynthesis in Dactylicapnos scandens by transcriptome analysis [J]. Sci Rep, 2017, 7(1): 9119.
33	Robin AY, Giustini C, Graindorge M, et al. Crystal structure of norcoclaurine-6-O-methyltransferase, a key rate-limiting step in the synthesis of benzylisoquinoline alkaloids [J]. Plant J, 2016, 87(6): 641-653.
34	Pyne ME, Kevvai K, Grewal PS, et al. A yeast platform for high-level synthesis of tetrahydroisoquinoline alkaloids [J]. Nat Commun, 2020, 11(1): 3337.
35	于荣敏, 王春盛, 宋丽艳. 罂粟科植物的化学成分及药理作用研究进展 [J]. 上海中医药杂志, 2004, 38(7): 59-61.
	Yu RM, Wang CS, Song LY. Progress on the study of chemical constituents and pharmacological action for the plants of Papaveraceae [J]. Shanghai J Tradit Chin Med, 2004, 38(7): 59-61.
36	Morris JS, Facchini PJ. Molecular origins of functional diversity in benzylisoquinoline alkaloid methyltransferases [J]. Front Plant Sci, 2019, 10: 1058.
37	Clapés P, Joglar J, Bujons J. Cascade biocatalysis: Integrating stereoselective and environmentally friendly reactions [M] . Hoboken: John Wiley & Sons, Inc, 2014.
38	Navarro G, Elliott HW. The effects of morphine, morphinone and thebaine on the EEG and behavior of rabbits and cats [J]. Neuropharmacology, 1971, 10(4): 367-377.
39	Ma WX, Noble WS, Bailey TL. Motif-based analysis of large nucleotide data sets using MEME-ChIP [J]. Nat Protoc, 2014, 9(6): 1428-1450.
40	Yang M, Fehl C, Lees KV, et al. Functional and informatics analysis enables glycosyltransferase activity prediction [J]. Nat Chem Biol, 2018, 14(12): 1109-1117.
41	朱新焰, 杨维泽, 杨绍兵, 等. 紫金龙营养器官组织结构和组织化学定位研究 [J]. 西南农业学报, 2022, 35(2): 304-309.
	Zhu XY, Yang WZ, Yang SB, et al. Tissue structure and histochemistry localization from different vegetative organs of Dactylicapnos scandens [J]. Southwest China J Agric Sci, 2022, 35(2): 304-309.
42	曹愿, 高晶, 高小力, 等. 紫金龙属生物碱及其药理活性研究进展 [J]. 中草药, 2014, 45(17): 2556-2563.
	Cao Y, Gao J, Gao XL, et al. Research progress on alkaloids from genus Dactylicapnos Wall. and their pharmacological activities [J]. Chin Tradit Herb Drugs, 2014, 45(17): 2556-2563.
43	Pienkny S, Brandt W, Schmidt J, et al. Functional characterization of a novel benzylisoquinoline O-methyltransferase suggests its involvement in papaverine biosynthesis in opium poppy (Papaver somniferum L) [J]. Plant J, 2009, 60(1): 56-67.
44	Xu DQ, Lin HF, Tang YP, et al. Integration of full-length transcriptomics and targeted metabolomics to identify benzylisoquinoline alkaloid biosynthetic genes in Corydalis yanhusuo [J]. Hortic Res, 2021, 8(1): 16.

编辑推荐 0

Metrics

阅读次数

全文

HTML			PDF

最新录用	在线预览	正式出版	最新录用	在线预览	正式出版
0	0	10	0	0	25

来源	本网站	其他网站

次数	31	4
比例	89%	11%

摘要

最新录用	在线预览	正式出版

0	0	56

	来源	本网站

	次数	56
	比例	100%

[1]	黄颖, 遇文婧, 刘雪峰, 刁桂萍. 山新杨谷胱甘肽转移酶基因的生物信息学与表达模式分析[J]. 生物技术通报, 2025, 41(2): 248-256.
[2]	向春繁, 李勒松, 王娟, 梁艳丽, 杨生超, 栗孟飞, 赵艳. 当归肉桂醇脱氢酶AsCAD功能鉴定及表达分析[J]. 生物技术通报, 2025, 41(2): 295-308.
[3]	王子傲, 田瑞, 崔永梅, 白羿雄, 姚晓华, 安立昆, 吴昆仑. 青稞HvnJAZ4的生物信息学和表达模式分析[J]. 生物技术通报, 2025, 41(1): 173-185.
[4]	孔青洋, 张晓龙, 李娜, 张晨洁, 张雪云, 于超, 张启翔, 罗乐. 单叶蔷薇GRAS转录因子家族鉴定及表达分析[J]. 生物技术通报, 2025, 41(1): 210-220.
[5]	吴娟, 武小娟, 王沛捷, 谢锐, 聂虎帅, 李楠, 马艳红. 彩色马铃薯花青素合成相关ERF基因筛选及表达分析[J]. 生物技术通报, 2024, 40(9): 82-91.
[6]	宋兵芳, 柳宁, 程新艳, 徐晓斌, 田文茂, 高悦, 毕阳, 王毅. 马铃薯G6PDH基因家族鉴定及其在损伤块茎的表达分析[J]. 生物技术通报, 2024, 40(9): 104-112.
[7]	武帅, 辛燕妮, 买春海, 穆晓娅, 王敏, 岳爱琴, 赵晋忠, 吴慎杰, 杜维俊, 王利祥. 大豆GS基因家族全基因组鉴定及胁迫响应分析[J]. 生物技术通报, 2024, 40(8): 63-73.
[8]	孙慧琼, 张春来, 王锡亮, 徐宏申, 窦苗苗, 杨博慧, 柴文婷, 赵珊珊, 姜晓东. 藜麦FLS基因家族的鉴定、表达及DNA变异分析[J]. 生物技术通报, 2024, 40(7): 172-182.
[9]	刘蓉, 田闵玉, 李光泽, 谭成方, 阮颖, 刘春林. 甘蓝型油菜REVEILLE家族鉴定及诱导表达分析[J]. 生物技术通报, 2024, 40(6): 161-171.
[10]	李嘉欣, 李鸿燕, 刘丽娥, 张恬, 周武. 沙棘NRAMP基因家族鉴定及铅胁迫下表达分析[J]. 生物技术通报, 2024, 40(5): 191-202.
[11]	潘萍萍, 徐志浩, 张怡雯, 李青, 王忠华. 多花黄精查尔酮合酶PcCHS的原核表达、亚细胞定位及表达分析[J]. 生物技术通报, 2024, 40(5): 280-289.
[12]	张娜, 刘梦楠, 屈展帆, 崔祎平, 倪嘉瑶, 王华忠. 小麦烯醇化酶基因ENO2的可变翻译分析和原核表达[J]. 生物技术通报, 2024, 40(5): 112-119.
[13]	钟匀, 林春, 刘正杰, 董陈文华, 毛自朝, 李兴玉. 芦笋皂苷合成相关糖基转移酶基因克隆及原核表达分析[J]. 生物技术通报, 2024, 40(4): 255-263.
[14]	郝楠, 耿珊, 赵雨薇, 侯智涵, 赵斌, 刘颖超. 拟轮枝镰孢丙氨酸转氨酶FvALT的克隆与表达分析[J]. 生物技术通报, 2024, 40(12): 256-263.
[15]	杨冲, 程莎莎, 艾长丰, 赵璇, 刘孟军. 枣ABF/AREB基因家族鉴定及其在果实发育中的表达分析[J]. 生物技术通报, 2024, 40(11): 184-191.

紫金龙异紫堇定生物合成相关6-OMT基因克隆与功能表征

Cloning and Functional Characterization of 6-OMT Gene Related to Isocorydine Biosynthesis in Dactylicapnos scandens

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 13

参考文献 44

相关文章 15

编辑推荐 0

Metrics

本文评价