牡丹ACC氧化酶基因cDNA克隆及全序列分析

 以牡丹品种‘洛阳红’（Paeonia suffruticosa 'Luoyang Hong'）花瓣为材料，用CTAB法提取总RNA，根据已报道的ACC氧化酶（1-aminocyclopropane-1-carboxylic acid oxidase，ACO）保守氨基酸序列设计简并引物，通过RT-PCR扩增得到1条821 bp的牡丹ACO基因同源片段。利用该已知中间序列，通过快速扩增cDNA末端技术(Race)及序列拼接，最终得到该基因cDNA全长序列，命名为Ps-ACO1，GenBank登录号为DQ337251。分析结果表明，Ps-ACO1 cDNA全长1 221 bp，包含一个939 bp的开放读码框，5'非翻译区长65 bp，3'非翻译区长117 bp，编码产物为含有312个氨基酸残基的蛋白质。氨基酸序列与烟草、苹果、桃等植物的ACO同源性都达80%以上。

Cloning and Sequence Analysis of 1-Aminocyciopropane-1-Carboxylic Acid Oxidase Gene cDNA from Tree Peony

Total RNA was extracted from petals of 'Luoyang Hong' tree peony (Paeonia suffruticosa) with CTAB method, and a pair of degenerated primer was designed based on the reported conserved amino acid sequence of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO). A cDNA fragment with 821 bp was amplified by RT-PCR, and then full length of it, named Ps-ACO1 (GenBank No. DQ337251), was obtained by RACE and sequence spliced. Sequence analysis indicated that Ps-ACO1 is 1 221 bp in full length and contains a 939 bp open reading frame flanking by a 65 bp 5′-non-translation region and a 117 bp 3′-non-translation region, and encodes a 321 predicated amino acid residues which shared more than 80% homology with ACO of tobacco, apple, and peach and so on.