茶树氧甲基转移酶基因的克隆及原核表达

doi:10.13305/j.cnki.jts.2013.06.014

Abstract

Abstract: A full length cDNA of O-methyltransferase gene was obtained from Camellia sinensis and the prokaryotic expression vector for this gene was constructed. Based on total RNA from tea leaves, a O-methyltransferase cDNA sequence of tea was obtained by RT-PCR and RACE. The whole cDNA sequence 1β280βbp which contains an ORF of 1β068βbp and encodes 355 amino acids. The putative protein of this gene had an isoelectric point of 5.68 and a calculated molecular weight of 39.1βkD. The amino acid sequence of tea O-methyltransferase showed 73%, 71% identity with that of Vitis vinifera and Ricinus communis respectively. The coding sequence had been cloned into pET-28a and transformed into the host BL21. Results of SDS-PAGE showed that the specific fusion protein was successfully induced to express by IPTG.

Key words: tea, O-methyltransferase, cloning, prokaryotic expression

CLC Number:

S571.1
Q555

MA Cheng-ying, SHI Jiang, LV Hai-peng, ZHANG Yue, TAN Jun-feng, GUO Li, PENG Qun-hua, LIN Zhi. Cloning and Prokaryotic Expression of O-methyltransferase from Camellia sinensis[J]. Journal of Tea Science, 2013, 33(6): 532-540.

References 27

[1]	田铃, 嵇保中, 刘曙雯, 等. 甲基转移酶的功能与分类[J]. 生命的化学, 2007, 27(5): 425-427.
[2]	李波, 倪志勇, 王娟, 等. 木质素生物合成关键酶咖啡酸-O-甲基转移酶基因(COMT)的研究进展[J]. 分子植物育种, 2010, 8(1): 117-124.
[3]	张传丽, 仲月明, 沈丹红, 等. 植物类黄酮O-甲基转移酶研究进展[J]. 西北植物学报, 2012, 32(6): 1274-1281.
[4]	Boudet AM, Lapierre C, Grima-Pettenati J.Bio-chemistry and molecular biology of lignifications[J]. New Phytologist, 1995, 129: 203-236.
[5]	Li L, Popko JL, Zhang XH, et al. A novel multifunctional O-methyltransferase implicated in a dual methylation pathway associated with lignin biosynthesis in loblolly pine[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94: 5461-5466.
[6]	Li L, Zhou Y, Cheng X, et al. Combinatorial modification of multiple lignin traits in trees through multigene cotransformation[J]. Proceedings of the National Academy of Sciences of the United States of America, 2003, 100: 4939-4944.
[7]	Muzac I, Wang J, Anzellotti D, et al. Functional Expression of an Arabidopsis cDNA Clone Encoding a Flavonol 3′-O-Methyltransferase and Characterization of the Gene Product[J]. Archives of Biochemistry and Biophysics, 2000, 375(2): 385-388
[8]	French CJ, Elder M, Leggett F, et al. Flavonoids inhibit infectivity of tobacco mosaic virus[J]. Canadian Journal of Plant Pathology, 1991, 13(1): 1-6.
[9]	Long SR.Rhizobium-legume nodulation: life together in the underground[J]. Cell, 1989, 56(2): 203-214.
[10]	Yang H, Ahn JH, Ibrahim RK, et a1. The three-dimensional structure of Arabidopsis thaliana O-methyltransferase predicted by homology-based modeling[J]. Journal Molecular Graph Model, 2004, 23(1): 77-87.
[11]	Maeda-Yamamoto M, Nagai H, Asai K, et al. Changes in epigallocatechin-3-O-(3-O-methyl gallate and strictinin contents of tea (Camellia sinensis L.) cultivar 'Benifuki' in various degrees of maturity and leaf order[J]. Food Science and Technology Research, 2004, 10(2): 186-190.
[12]	Kelavkar UP, Nixon JB, Cohen C, et al. Overexpression of 15-lipoxygenase-1 in PC-3 human prostate cancer cells increases tumorigenesis[J]. Carcinogenesis, 2001, 22(11): 1765-1773.
[13]	Kurita I, Maeda YM, Tachibana H, et al. Antihypertensive effect of Benifuuki tea containing O-methylated EGCG[J]. Journal of Agricultural and Food Chemistry, 2010, 58(3): 1903-1908.
[14]	Wu SQ, Watanabe N, Mita S, et al. Two O-Methyltransferases isolated from flower petals of Rosa chinensis var. spontanea involved in scent biosynthesis[J]. Journal of Bioscience and Bioengineering, 2003, (96): 119-128.
[15]	Joshi CP, Chiang VL.Conserved sequence motifs in plant S-adenosyl-L-methionine-dependent methyltransferase[J]. Plant Molecular Biology, 1998, 37: 663-674.
[16]	Bong-Gyu Kim, Su Hyun Sung, Youhoon Chong, et al. Plant Flavonoid O-Methyltransferases: Substrate Specificity and Application[J]. J Plant Biology, 2010, 53: 321-329.
[17]	Schmitt D, Pakusch AE, Matern U.Molecular cloning, induction and taxonomic distribution of caffeoyl-CoA 3-O-methyltransferase, an enzyme involved in disease resistance[J]. Biological Chemistry, 1991, 266: 17416-17423.
[18]	马成英, 吕海鹏, 林智, 等. 茶树类黄酮O-甲基转移酶基因的克隆及原核表达分析[J]. 中国农业科学, 2013, 46(2): 325-333.
[19]	李小霞, 余有本. 茶树咖啡酸氧甲基转移酶基因的克隆及原核表达[J]. 西北农业学报, 2011, 20(5): 139-143.
[20]	Masanobu K, Daiki H, Yoshihisa T, et al. Cloning of a novel O-methyltransferase from Camellia sinensis and synthesis of O-methylated EGCG and evaluation of their bioactivity[J]. Journey of Agricultural and Food Chemistry, 2010, 58(12): 7196-7201.
[21]	费冬梅, 林智, 吕海鹏, 等. EGCG-O-甲基转移酶(EOMT)在重组大肠杆菌中的表达条件研究[J]. 茶叶科学, 201l, 31(4): 333-340.
[22]	Kces RE, Quattrocchio F, Mol JNM.The flavonoid biosynthetic pathway in plants: function and evolution[J]. BioEssays, 1994, 16(2): 123-132.
[23]	Gauthier A, Patrick JG, Ibrahim R K, et al. Characterization of two cDNA clones which encode O-methyltransferases for the methylation of both flavonoid and phenylpropanoid compounds[J]. Archives of Biochemistry and Biophysics, 1998, 351(2): 243-249.
[24]	Frick S, Kutchan TM.Molecular cloning and functional expression of O-methyltransferases common to isoquinoline alkaloid and phenylpropanoid biosynthesis[J]. Plant J, 1999, 17: 329-339.
[25]	Schroder G, Wehinger E, Schroder J.Predicting the substrates of cloned plant O-methyltransferases[J]. Phytochemistry, 2002, 59: 1-8.
[26]	Wang J, Pichersky E.Identiﬁcation of speciﬁc residues involved in substrate discrimination in two plant O-methyltransferases[J]. Archives of Biochemistry and Biophysics, 1999, 368: 172-180.
[27]	Zubieta C, He XZ,Dixon RA, et al. Structures of two natural product methyhransferases reveal the basis for substrate specificity in plant O-methyltransferases[J]. Nature Structural Biology, 2001, 8: 271-279.

[1]	. [J]. Journal of Tea Science, 1965, 2(04): 59 -63 .
[2]	. [J]. Journal of Tea Science, 1965, 2(03): 18 -21 .
[3]	. [J]. Journal of Tea Science, 1965, 2(03): 22 -24 .
[4]	. [J]. Journal of Tea Science, 1966, 3(02): 55 -57 .
[5]	. [J]. Journal of Tea Science, 1966, 3(02): 82 -83 .
[6]	. [J]. Journal of Tea Science, 1986, 6(01): 1 -8 .
[7]	. [J]. Journal of Tea Science, 1987, 7(02): 29 -34 .
[8]	. [J]. Journal of Tea Science, 1988, 8(01): 37 -42 .
[9]	. [J]. Journal of Tea Science, 1995, 15(02): 154 .
[10]	. [J]. Journal of Tea Science, 1999, 19(01): 73 -76 .

Cloning and Prokaryotic Expression of O-methyltransferase from Camellia sinensis

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 27

Related Articles 15

Metrics

Comments

Recommended 10

[1]	GAO Jianjian, CHEN Dan, PENG Jiakun, WU Wenliang, CAI Liangsui, CAI Yawei, TIAN Jun, WAN Yunlong, SUN Weijiang, HUANG Yan, WANG Zhe, LIN Zhi, DAI Weidong. Comparison on Chemical Components of Yunnan and Fuding White Tea Based on Metabolomics Approach [J]. Journal of Tea Science, 2022, 42(5): 623-637.
[2]	CHEN Qiyu, MA Jianqiang, CHEN Jiedan, CHEN Liang. Genetic Diversity of Mature Leaves of Tea Germplasms Based on Image Features [J]. Journal of Tea Science, 2022, 42(5): 649-660.
[3]	LI Yanchun, WANG Hang, LI Zhaowei, YE Jing, WANG Yixiang. Ameliorative Effect of Several Measures on Soil Physicochemical Properties and Microbial Community Structures in Acidified Tea Gardens [J]. Journal of Tea Science, 2022, 42(5): 661-671.
[4]	SUN Yue, WU Jun, WEI Chaoling, LIU Mengyue, GAO Chenxi, ZHANG Lingzhi, CAO Shixian, YU Shuntian, JIN Shan, SUN Weijiang. Screening of Tea Germplasm Resistant to Matsumurasca onukii and Dendrothrips minowai Priesner and Analysis of Resistance-related Factors [J]. Journal of Tea Science, 2022, 42(5): 689-704.
[5]	CHEN Yuhong, GAO Ying, HAN Zhen, YIN Junfeng. Analysis of the Saponin Contents and Composition in Tea Seeds of Different Germplasms [J]. Journal of Tea Science, 2022, 42(5): 705-716.
[6]	CHEN Hui, YANG Liling, CHEN Jinhua, HUANG Jian'an, GONG Yushun, LI Shi. Effect of Temperature-controlled Pile-fermentation on Aroma Quality of Primary Dark Tea [J]. Journal of Tea Science, 2022, 42(5): 717-730.
[7]	LI Zheng, LIU Ding, HUO Zenghui, CHEN Fuqiao. Analysis on the Competitiveness and Complementarity of Tea Trade between China and RCEP Members [J]. Journal of Tea Science, 2022, 42(5): 740-752.
[8]	WANG Yuyuan, LIU Renjian, LIU Shaoqun, SHU Canwei, SUN Binmei, ZHENG Peng. Expression Analysis and Functional Identification of CsTT2 R2R3-MYB Transcription Factor in Tea Plants [J]. Journal of Tea Science, 2022, 42(4): 463-476.
[9]	LI Jing, LIN Cairong, HUANG Yan, DENG Xuming, WANG Yiqing, SUN Weijang. Effects of Tea Polyphenols on Agrobacterium-mediated Plant Genetic Transformation System [J]. Journal of Tea Science, 2022, 42(4): 477-490.
[10]	ZHAO Dongwei. Nomenclature, Typification, and Natural Distribution of Camellia sinensis var. assamica (Theaceae) [J]. Journal of Tea Science, 2022, 42(4): 491-499.
[11]	LIU Jianjun, ZHANG Jinyu, PENG Ye, LIU Xiaobo, YANG Yun, HUANG Tao, WEN Beibei, LI Meifeng. Effects of Light Waves on the Aroma Substances of Fresh Tea Leaves in Summer and Autumn During Spreading and the Quality of Final Green Tea [J]. Journal of Tea Science, 2022, 42(4): 500-514.
[12]	ZHANG Yinggen, XIANG Lihui, CHEN Lin, LIN Qingxia, SONG Zhenshuo, WANG Lili. Effects of Air Conditions Preset for Withering on Flavor Quality and Chemical Profiles of White Teas [J]. Journal of Tea Science, 2022, 42(4): 525-536.
[13]	LI Ziqiang, YANG Mei, ZHANG Xinzhong, LUO Fengjian, ZHOU Li, LOU Zhengyun, SUN Hezhi, WANG Xinru, CHEN Zongmao. Residue Determination of Sixteen Pesticides in Green Tea by UPLC-MS/MS [J]. Journal of Tea Science, 2022, 42(4): 537-548.
[14]	FANG Mengrui, LÜ Jun, RUAN Jianyun, BIAN Lei, WU Chuanyu, YAO Qing. Tea Buds Detection Model Using Improved YOLOv4-tiny [J]. Journal of Tea Science, 2022, 42(4): 549-560.
[15]	SUN Ying, CHEN Xin, YANG Hua, YING Jian, SHAO Danqing, LÜ Xiaohua, XIAO Jie, CHEN Zhixiong, LI Song, QIN Junjie, ZHENG Bin, GAO Jianshe. Clinical Trial on the Effect of Drinking Jinhua Xiangyuan Tea for 3 Months on the Improvement of Glucose and Lipid Metabolism in A Small Sample Hyperlipidemia Population [J]. Journal of Tea Science, 2022, 42(4): 561-576.