油橄榄查尔酮合酶与查尔酮异构酶基因全长的克隆及序列分析

doi:10.7506/spkx1002-6630-201509022

食品科学

油橄榄查尔酮合酶与查尔酮异构酶基因全长的克隆及序列分析

陈文拴1，黄乾明1,*，陈华萍1，杨泽身2，王安逸2，苏光灿2

1.四川农业大学理学院，四川雅安 625014；2.凉山州中泽新技术开发有限责任公司，四川西昌 615000

出版日期:2015-05-15 发布日期:2015-05-11

Cloning and Characterization of Full-Length Chalcone Synthase and Chalcone Isomerase Genes from Olea europaea

CHEN Wenshuan1, HUANG Qianming1,*, CHEN Huaping1, YANG Zeshen2, WANG Anyi2, SU Guangcan2

1. College of Science, Sichuan Agricultural University, Ya’an 625014, China;
2. Liang Shan Zhong Ze New Technology Development Co. Ltd., Xichang 615000, China

Online:2015-05-15 Published:2015-05-11

摘要/Abstract

摘要：

查尔酮合酶（chalcone synthase，Chs）与查尔酮异构酶（chalcone isomerase，Chi）是植物类黄酮合成代谢途径中的两个关键酶。采用同源克隆、反转录聚合酶链式反应（reverse transcription polymerase chain reaction，RT-PCR）、融合引物嵌套PCR（fusion primer and nested integrated PCR，FPNI-PCR）与3’-cDNA末端快速扩增技术（rapid amplification of cDNA end，RACE）相结合，分别克隆出油橄榄chs与chi基因全长。序列分析表明，油橄榄chs基因全长DNA（GenBank登录号：KF935223.1）和cDNA（GenBank登录号：KF935224.1）序列分别为2 085 bp和1 173 bp，该基因的开放阅读框（open reading frame，ORF）编码390 个氨基酸，其氨基酸序列与葡萄和番茄全长Chs氨基酸序列的相似度在90%左右。生物信息学分析表明，该基因编码的蛋白属于Chs蛋白家族。油橄榄chi基因全长DNA（GenBank登录号：KF886190）和cDNA（GenBank登录号：KF886191）序列分别为1 373 bp和750 bp，其ORF编码249 个氨基酸，该氨基酸序列与已报道的其他植物Chi氨基酸序列的相似度最高在65%左右，序列中包含一段由198 个氨基酸残基组成的具有分子内裂解酶活性的功能域，符合Chi蛋白家族特征。

关键词: 油橄榄, 查尔酮合酶, 查尔酮异构酶, 基因克隆, 序列分析

Abstract:

Chalcone synthase (Chs) and chalcone isomerase (Chi) are two key enzymes in the biosynthesis pathway of plant
flavonoids. The complete genes of chs and chi were isolated from Olea europaea by homology cloning, RT-PCR, FPNI-PCR
(fusion primer and nested integrated PCR) and 3’-RACE (rapid amplification of cDNA end). Sequence analysis showed that
the full-length DNA (GenBank No: KF935223.1) and cDNA (GenBank No: KF935224.1) of chs gene from O. europaea
were 2 085 bp and 1 173 bp, respectively, and the ORF (open reading frame) of chs gene encoded 390 amino acid residues,
which showed high similarity (about 90%) with the complete amino acid sequences of Chs from Vitis vinifera and Solanum
lycopersicum. Bioinformatics analysis showed that the protein encoded by chs gene from O. europaea belonged to the Chs
protein family. The full-length DNA (GenBank No: KF886190) and cDNA (GenBank No: KF886191) of chi gene from
O. europaea were 1 373 bp and 750 bp, respectively, and the ORF of chi gene encoded 249 amino acids residues. With the
highest similarity (about 65%) to the complete amino acid sequences of Chi from the other plants , the full-length amino acid
sequence of Chi from O. europaea had an intramolecular lyase domain with 198 amino acids, which was consistent with the
characteristics of the Chi protein family.

Key words: Olea europaea, chalcone synthase (Chs), chalcone isomerase (Chi), gene cloning, sequence analysis

中图分类号:

Q943.2

陈文拴1，黄乾明1,*，陈华萍1，杨泽身2，王安逸2，苏光灿2. 油橄榄查尔酮合酶与查尔酮异构酶基因全长的克隆及序列分析[J]. 食品科学, doi: 10.7506/spkx1002-6630-201509022.

CHEN Wenshuan1, HUANG Qianming1,*, CHEN Huaping1, YANG Zeshen2, WANG Anyi2, SU Guangcan2. Cloning and Characterization of Full-Length Chalcone Synthase and Chalcone Isomerase Genes from Olea europaea[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201509022.

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油橄榄查尔酮合酶与查尔酮异构酶基因全长的克隆及序列分析

Cloning and Characterization of Full-Length Chalcone Synthase and Chalcone Isomerase Genes from Olea europaea

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