药学学报, 2019, 54(7): 1312-1316
引用本文:
郝丹丹, 胡启跳, 袁媛, 龚永福, 陈芳, 魏玉杰, 黄璐琦. 罂粟与虞美人可待因-O-脱甲基酶基因克隆及序列分析[J]. 药学学报, 2019, 54(7): 1312-1316.
HAO Dan-dan, HU Qi-tiao, YUAN Yuan, GONG Yong-fu, CHEN Fang, WEI Yu-jie, HUANG Lu-qi. Cloning and sequence analysis of codeine-O-demethylase gene from Papaver somniferum and Papaver rhoeas[J]. Acta Pharmaceutica Sinica, 2019, 54(7): 1312-1316.

罂粟与虞美人可待因-O-脱甲基酶基因克隆及序列分析
郝丹丹1,2, 胡启跳2, 袁媛2, 龚永福3, 陈芳3, 魏玉杰3, 黄璐琦2
1. 山西中医药大学中药学院, 山西 晋中 030619;
2. 中国中医科学院中药资源中心, 北京 100700;
3. 甘肃省农业工程技术研究院, 甘肃 武威 733000
摘要:
CODM是可待因和吗啡生物合成过程中的关键酶,本实验以NCBI数据库中的可待因-O-脱甲基酶基因(codeine-O-demethylase,CODM)序列为参考,克隆得到了罂粟和虞美人的CODM核苷酸序列,并对二者进行序列比对及生物信息学分析。结果发现,罂粟CODM有3种基因型,而虞美人CODM有5种基因型;生物信息学分析显示所有CODM蛋白无信号肽序列,预测这些蛋白都为非分泌性蛋白,属于Pcbc superfamily超基因家族。罂粟CODM氨基酸序列相同,但在不同罂粟资源中的表达量有明显差异,推测CODM基因转录水平变异可能与其非编码区序列有关,这为进一步开展罂粟生物碱类成分的合成与调控机制研究奠定基础。
关键词:    罂粟      虞美人      可待因-O-脱甲基酶      生物信息学分析     
Cloning and sequence analysis of codeine-O-demethylase gene from Papaver somniferum and Papaver rhoeas
HAO Dan-dan1,2, HU Qi-tiao2, YUAN Yuan2, GONG Yong-fu3, CHEN Fang3, WEI Yu-jie3, HUANG Lu-qi2
1. School of Chinese Materia Medica, Shanxi University of Chinese Medicine, Jinzhong 030619, China;
2. National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;
3. Gansu Institute of Agricultural Engineering Technology, Wuwei 733000, China
Abstract:
Codeine-O-demethylase (CODM) is a key enzyme in the biosynthesis of codeine and morphine. In this study, CODM gene sequences were cloned from Papaver somniferum and Papaver rhoeas, and were compared with each other by sequence alignment and bioinformatics analysis. The results showed that there were three genotypes of CODM in Papaver somniferum and five genotypes of CODM in Papaver rhoeas. Bioinformatics analysis showed that all CODM proteins had no signal peptide sequence, and these proteins were predicted to be non-secretory proteins, belonging to the Pcbc supergene family. Although the amino acid sequences of CODM in poppies are the same, the expression levels of CODM in different poppy resources are significantly different. It is speculated that the variation of transcription level of CODM may be related to its non-coding region sequence, which lays a foundation for further research on the synthesis and regulation mechanism of alkaloids in poppies.
Key words:    Papaver somniferum    Papaver rhoeas    codeine-O-demethylase    bioinformatics analysis   
收稿日期: 2019-03-11
DOI: 10.16438/j.0513-4870.2019-0163
基金项目: 中央级公益性科研院所基本科研业务费专项资金资助(ZZ10-008);中央本部级重大支减项目:名贵中药资源可持续利用能力建设项目(2060302);成都中医药大学西南特色中药资源重点实验室开放研究基金资助项目(003109034001).
通讯作者: 袁媛,Tel:86-10-64087649,E-mail:y_yuan0732@163.com
Email: y_yuan0732@163.com
相关功能
PDF(2875KB) Free
打印本文
0
作者相关文章
郝丹丹  在本刊中的所有文章
胡启跳  在本刊中的所有文章
袁媛  在本刊中的所有文章
龚永福  在本刊中的所有文章
陈芳  在本刊中的所有文章
魏玉杰  在本刊中的所有文章
黄璐琦  在本刊中的所有文章

参考文献:
[1] Hagel JM, Facchini PJ. Benzylisoquinoline alkaloid metabolism:a century of discovery and a brave new world[J]. Plant Cell Physiol, 2013, 54:647-672.
[2] Choe S, Kim S, Lee C, et al. Species identification of Papaver by metabolite profiling[J]. Forensic Sci Int, 2011, 211:51-60.
[3] Coban I, Toplan GG, Özbek B, et al. Variation of alkaloid contents and antimicrobial activities of Papaver rhoeas L. growing in Turkey and northern Cyprus[J]. Pharm Biol, 2017, 55:1894-1898.
[4] Oh JH, Ha IJ, Lee MY, et al. Identification and metabolite profiling of alkaloids in aerial parts of Papaver rhoeas by liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry[J]. J Sep Sci, 2018, 41:2517-2527.
[5] Hagel JM, Facchini PJ. Dioxygenases catalyze the O-demethylation steps of morphine biosynthesis in opium poppy[J]. Nat Chem Biol, 2010, 6:273-275.
[6] Wijekoon CP, Facchini PJ. Systematic knockdown of morphine pathway enzymes in opium poppy using virus-induced gene silencing[J]. Plant J, 2012, 69:1052-1063.
[7] Chen TY, Liu J, Yuan Y, et al. Bioinformatics analysis and expressed level of Auxin response factor genes in Scutellaria baicalensis[J]. Acta Pharm Sin (药学学报), 2017, 52:1770-1776.
[8] Facchini PJ,Park SU. Developmental and inducible accumulation of gene transcripts involved in alkaloid biosynthesis in opium poppy[J]. Phytochemistry, 2003, 64:177-186.
[9] Guo L, Winzer T, Yang X, et al. The opium poppy genome and morphinan production[J]. Science, 2018, 362:343-347.