[1]廖燕,尹松松,龚林涛,等.薰衣草醇酰基转移酶基因（AAT）的克隆和表达分析[J].江苏农业学报,2024,(04):599-606.[doi:doi:10.3969/j.issn.1000-4440.2024.04.003]
　LIAO Yan,YIN Song-song,GONG Lin-tao,et al.Cloning and expression analysis of lavender alcohol acyltransferase gene （AAT）[J].,2024,(04):599-606.[doi:doi:10.3969/j.issn.1000-4440.2024.04.003]
点击复制

薰衣草醇酰基转移酶基因（AAT）的克隆和表达分析()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2024年04期
页码:
599-606
栏目:
遗传育种·生理生化
出版日期:
2024-04-30

文章信息/Info

Title:
Cloning and expression analysis of lavender alcohol acyltransferase gene （AAT）
作者:
廖燕1; 尹松松1; 龚林涛1; 闫博文1; 孙明辉1; 苏秀娟1; 2; 王爱凡1; 2
(1.新疆农业大学农学院,新疆乌鲁木齐830052;2.新疆农业大学薰衣草研究所/新疆作物遗传改良与种质创新重点实验室，新疆乌鲁木齐830052)
Author(s):
LIAO Yan1; YIN Song-song1; GONG Lin-tao1; YAN Bo-wen1; SUN Ming-hui1; SU Xiu-juan1; 2; WANG Ai-fan1; 2
（1.College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China;2.Lavender Research Institute of Xinjiang Agricultural University/Xinjiang Key Laboratory of Crop Improvement & Germplasm Enhancement, Urumqi 830052, China）
关键词:
薰衣草; 醇酰基转移酶基因（AAT）; 克隆; 表达模式; 酶活性鉴定
Keywords:
lavender; alcohol acyltransferase gene (AAT); cloning; expression pattern; enzyme activity identification
分类号:
S184
DOI:
doi:10.3969/j.issn.1000-4440.2024.04.003
摘要:
醇酰基转移酶（AAT）可以催化乙酰辅酶A与萜烯醇类物质的结合，生成各类乙酸酯类化合物。为探明薰衣草AAT基因在酯类化合物合成中的调控作用，本研究以薰衣草品系杂花叶片为供试材料，利用反转录PCR技术克隆薰衣草AAT基因的cDNA序列，该cDNA序列长度为1 344 bp，其编码蛋白质由447个氨基酸组成，是非跨膜亲水性蛋白质，属于PLN02481超级家族；薰衣草AAT与黄色猴面花EgAAT亲缘关系最近，相似度为60.87%。AAT基因在苞叶中的表达量最低，在花瓣中的表达量最高；在花冠不同发育时期，该基因在盛开期表达量到达峰值。构建原核表达载体pET-28a-AAT，筛选了重组蛋白在大肠杆菌Transetta(DE3)中的最适诱导表达条件，诱导后获得的重组蛋白质相对分子质量为5.026×104，重组蛋白以包涵体形式存在，具有将芳樟醇催化合成乙酸芳樟酯的生物活性。本研究结果为进一步验证薰衣草AAT基因在酯类化合物合成过程中的功能提供了基础。
Abstract:
Alcohol acyltransferase (AAT) can catalyze the combination of acetyl coenzyme A and terpene alcohols to produce various types of acetate compounds. In order to explore the regulatory role of lavender AAT gene in the synthesis of ester compounds, the cDNA sequence of lavender AAT gene was cloned by reverse transcription PCR using the leaves of lavender strain Zahua as the test material. The length of the cDNA sequence was 1 344 bp, and the encoded protein was composed of 447 amino acids, which was a non-transmembrane hydrophilic protein and belonged to the PLN02481 superfamily. The AAT in lavender was closely related to the EgAAT in yellow monkey-faced flower, with a similarity of 60.87%. The expression level of AAT gene was lowest in bracts and highest in petals. At different developmental stages of corolla, the expression of this gene reached the peak at the full-bloom stage. Prokaryotic expression vector pET-28a-AAT was constructed. The optimal expression conditions of the recombinant protein in Escherichia coli Transetta (DE3) were screened. The relative molecular weight of the recombinant protein was about 5.026×104. The recombinant protein existed in the form of inclusion bodies and had the biological activity of catalyzing the synthesis of linalyl acetate from linalool. The results of this study provide a basis for further verifying the function of lavender AAT gene in the synthesis of ester compounds.

参考文献/References:

[1]张群,扎灵丽. 薰衣草的研究和应用[J]. 时珍国医国药,2008(6):1312-1314.
[2]王玉芹,孙亚军,施献儿. 薰衣草精油的化学成分与药理活性[J]. 国外医药(植物药分册),2004(1):5-8.
[3]张健,蔡宝国,章苏宁,等. 薰衣草精油化学成分的GC-MS分析比较[J]. 食品工业,2007(5):52-54.
[4]徐洁华,文首文. 薰衣草挥发性有机物及其药理功效研究进展[J]. 时珍国医国药,2010,21(4):979-980，983.
[5]PICHERSKY E, RAGUSO R A. Why do plants produce so many terpenoid compounds?[J]. The New Phytologist,2018,220(3):692-702.
[6]DEHSHEIKH A B, SOURESTANI M M, DEHSHEIKH P B, et al. Monoterpenes:essential oil components with valuable features[J]. Mini Reviews in Medicinal Chemistry,2020,20(20):958-974.
[7]VRANOV E, COMAN D, GRUISSEM W. Network analysis of the MVA and MEP pathways for isoprenoid synthesis[J]. Annual Review of Plant Biology,2013,64:665-700.
[8]岳跃冲,范燕萍. 植物萜类合成酶及其代谢调控的研究进展[J]. 园艺学报,2011,38(2):379-388.
[9]LAURA K H, GUTENSOHN M, SUZANNE T T, et al. Orthologs of the archaeal isopentenyl phosphate kinase regulate terpenoid production in plants[J]. Proceedings of the National Academy of Sciences of the United States of America,2015,112(32):10050-10055.
[10]刘琬菁,吕海舟,李滢,等. 植物萜类合酶研究新进展[J]. 植物生理学报,2017,53(7):1139-1149.
[11]魏恬恬,臧姝,曾雨倩,等. 植物单萜类化合物的代谢调控[J]. 扬州大学学报(农业与生命科学版),2017,38(2):122-126.
[12]SHALIT M, GUTERMAN I, VOLPIN H, et al. Volatile ester formation in roses.Identification of an acetyl-coenzyme A. Geraniol/citronellolacety-ltransferase in developing rose petals[J]. Plant Physiol,2003,8(1):153-162.
[13]李晋华. 月月粉乙酰基转移酶基因RcAAT的克隆及功能分析[D]. 昆明：云南农业大学,2017.
[14]CRISTIAN B, CARLOS G E, LIDA F, et al. VpAAT1, a gene encoding an alcohol acyltransferase, is involved in ester biosynthesis during ripening of mountain papaya fruit[J]. Journal of Agricultural and Food Chemistry,2010,58(8):5114-5121.
[15]董静,王桂霞,钟传飞,等. 森林草莓醇酰基转移酶基因FvAATW2功能研究[J]. 园艺学报,2018,45(1):41-50.
[16]LI D P, XU Y F, XU G G, et al. Molecular cloning and expression of a gene encoding alcohol acyltransferase (MdAAT2) from apple (cv. Golden Delicious)[J]. Phytochemistry,2006,67(7):658-667.
[17]龚林涛,苏秀娟,廖燕,等. 薰衣草芳樟醇合酶基因的克隆、表达及酶活性检测[J]. 作物杂志,2021,2021(6):78-87.
[18]尹松松,苏秀娟,龚林涛,等. 薰衣草1-脱氧-D-木酮糖-5-磷酸还原异构酶基因LaDXR的克隆及表达分析[J]. 分子植物育种,2021,19(7):2193-2199.
[19]龚林涛,苏秀娟,尹松松,等. 薰衣草DXS基因的克隆、表达分析及原核表达[J]. 新疆农业科学,2020,57(7):1233-1242.
[20]曹香梅. 桃果实酯类芳香物质的代谢与调控研究[D]. 杭州：浙江大学,2019.
[21]王梦姣. 植物BAHD酰基转移酶家族研究进展[J]. 江西农业学报,2010,22(12):124-126.
[22]刘雨雨,莫婷,王晓晖,等. 植物来源BAHD酰基转移酶家族研究进展[J]. 中国中药杂志,2016,41(12):2175-2182.
[23]刘偲,曾祥玲,郑日如,等. 桂花花瓣醇酰基转移酶基因的克隆与表达[J]. 华中农业大学学报,2016,35(1):36-42.
[24]ELHADI M A M. 柿果实品质特性、香气成分分析及醇酰基转移酶基因（DkAAT1）的表达研究[D]. 扬州：扬州大学,2017.
[25]霍婷婷. 梅花花香相关基因PmCFATs的克隆和表达分析[D]. 北京：北京林业大学,2017.
[26]陈陈. 玫瑰花香成分生物合成相关基因的克隆与表达分析[D]. 扬州：扬州大学,2012.
[27]曹颖,胡尚连,张慧莹,等. 番茄醇酰基转移酶基因SlAAT1克隆、序列分析和原核表达[J]. 植物研究,2012,32(6):731-736.
[28]刘博文,王雪庆,孙涛,等. 白蜡虫蜡酯合酶基因cDNA全长克隆及原核表达[J]. 林业科学研究,2016,29(4):610-614.
[29]BAESHEN N A, BAESHENM N, SHEIKH A, et al. Cell factories for insulin production [J]. Microbial Cell Factories,2014,13(1):141-149.
[30]FERRER-MIRALLES N, VILLAVERDE A. Bacterial cell factories for recombinant protein production; expanding the catalogue [J]. Microbial Cell Factories,2013,12(1):113.

备注/Memo

备注/Memo:
收稿日期：2023-04-27基金项目：新疆维吾尔自治区重大科技专项（2022B02036-1）；国家自然科学基金地区科学基金项目（31760429）；新疆维吾尔自治区科学技术厅“天山青年计划”优秀青年科技人才培养项目（2020Q016）；国家级大学生创新训练计划项目（201810758003)；作物学重点学科发展基金项目（XNCDKY2021015）作者简介：廖燕（2000-），女，重庆人，硕士研究生，主要从事薰衣草遗传育种研究。（E-mail）1642101080@qq.com通讯作者：苏秀娟，（E-mail）smm1980@yeah.net;王爱凡，（E-mail）wangaifantop@163.com
更新日期/Last Update: 2024-05-22