万寿菊玉米黄质环氧化酶TeZEP基因片段的克隆及分析  

Cloning and Analysis of Zeaxanthin Cyclooxygenase TeZEP Gene Fragment in Marigold

作　　者：刘双 于放[1] Liu Shuang;Yu Fang(College of Bioengineering,Dalian University of Technology,Dalian,116033)

机构地区：[1]大连工业大学生物工程学院,大连116033

出　　处：《分子植物育种》2020年第4期1077-1082,共6页Molecular Plant Breeding

基　　金：国家自然科学基金面上项目(31570303)资助。

摘　　要：为了研究万寿菊中玉米黄质生物合成途径的关键酶基因,首先根据不同物种的ZEP基因进行序列比对找到保守区域设计简并引物,然后利用RT-PCR方法,以万寿菊花瓣总RNA为模板克隆万寿菊ZEP基因cDNA的片段,并进行序列分析。结果表明,万寿菊ZEP基因片段长度为1 058 bp,编码352个氨基酸残基,其序列与其他植物ZEP基因同源性最大为92%;其翻译产物属于亲水性蛋白;且结构域预测结果发现TeZEP基因编码的氨基酸序列含有一个NADB_Rossmann超家族和玉米黄质环氧化酶的特征性结构黄素腺嘌呤二核苷酸(FAD)结合位点。本研究成功克隆了万寿菊玉米黄质环氧化酶TeZEP基因片段并对其进行了序列分析,这为进一步研究该基因功能提供了依据,也为构建转基因植物和沉默基因植株以提高玉米黄质合成量提供了一定的参考。In order to study the key enzyme genes in the zeaxanthin biosynthesis pathway in marigold, firstly, based on the sequence alignment of ZEP genes of different species, we found a conserved region to design degenerate primers. Then, a partial fragment of the marigold ZEP gene cDNA was cloned by RT-PCR using the total RNA of the marigold flower petals as a template, and analyzed the sequence. The results showed that the marigold ZEP gene fragment was 1 058 bp in length and encoded 352 amino acid residues, and its sequence had the highest homology with other plant ZEP genes of 92%;its translation product was a hydrophilic protein;In this study, the TeZEP gene fragment of marigold zeaxanthin epoxidase was successfully cloned and sequenced, which provided a basis for further study of the gene function. It also lays a foundation for the construction of transgenic plants and silent gene plants to increase the amount of zeaxanthin synthesis.

关 键 词：万寿菊(Tagetes ERECTA L.) ZEP 基因 序列分析 玉米黄质 

分 类 号：S681.9[农业科学—观赏园艺]