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薰衣草醇酰基转移酶基因(AAT)的克隆和表达分析

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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.
Cloning and expression analysis of lavender alcohol acyltransferase gene(AAT)
Alcohol acyltransferase(AAT)can catalyze the combination of acetyl coenzyme A and terpene alcohols to pro-duce 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 1344 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 bi-ological activity of catalyzing the synthesis of linalyl acetate from linalool.The results of this study provide a basis for fur-ther verifying the function of lavender AAT gene in the syn-thesis of ester compounds.

lavenderalcohol acyltransferase gene(AAT)cloningexpression patternenzyme activity identification

廖燕、尹松松、龚林涛、闫博文、孙明辉、苏秀娟、王爱凡

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新疆农业大学农学院,新疆 乌鲁木齐 830052

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

薰衣草 醇酰基转移酶基因(AAT) 克隆 表达模式 酶活性鉴定

新疆维吾尔自治区科技重大专项国家自然科学基金地区科学基金新疆维吾尔自治区科技厅"天山青年计划"优秀青年科技人才培养项目国家级大学生创新训练项目作物学重点学科发展基金

2022B02036-1317604292020Q016201810758003XNCD-KY2021015

2024

10.3969/j.issn.1000-4440.2024.04.003

江苏农业学报
江苏省农业科学院

江苏农业学报

CSTPCD北大核心
影响因子：1.093
ISSN：1000-4440
年,卷(期)：2024.40(4)
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