狗头枣ACC I氧化酶基因的克隆及其序列分析

Cloning and Sequence Analysis of ACC I Oxidase Gene from Zizyphus jujube(Goutouzao)

为利用RNAi技术开展调控乙烯合成延迟狗头枣果实成熟等研究,以狗头枣试管苗叶片的总DNA为模板,根据冬枣ACC I氧化酶基因的序列设计引物,通过PCR方法获得一长1 294 bp片段,序列分析结果表明,该片段具有4个外显子和3个内含,依次在106～201、430～541 bp与877～1 002 bp碱基处.此序列与冬枣ACC I氧化酶基因（EU216549.1）的核苷酸和氨基酸序列同源性均为99.69％,表明成功地克隆了狗头枣ACC I氧化酶基因.

In order to carry out researching of delaying Goutou jujube fruit ripening by regulation of ethylene synthesis with RNAi technology,a pair of specific primers were designed according to ACC I oxidase gene sequence of Zizyphus jujube （Dongzao） recorded in GenBank.Using test-tube plantlet total DNA from ＂Goutou jujube＂ as template,a fragment of ACC I oxidase gene including an ORF （1 294 bp） was amplified.The results showed that the fragment has four exons and three introns,turn in at 106～201,430～541 and 877～1 002 bp base.Compared with nucleotide sequence and amino acid sequence homology of ACC I oxidase gene （EU216549.1） landed were 99.69％,which showed the successful cloning of the Goutou jujube ACC I oxidase gene.