红花生育酚环化酶基因的克隆及表达分析  

Cloning and expression analysis of tocopherol cyclase gene in Carthamus tinctorius

作　　者：官丽莉[1] 崔琪[1] 韩雨彤[2] 武云云[1] 胡人阁 顾天瑶 李海燕[1] 李校堃[1] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]东北师范大学附属实验中学净月实验学校,吉林长春130118

出　　处：《中国中药杂志》2016年第20期3727-3732,共6页China Journal of Chinese Materia Medica

基　　金：国家高技术研究发展计划(863)项目(2011AA100606);国家自然科学基金项目(31201237);国家级大学生创新创业项目(201610193046;201410193045);吉林省教育厅"十三五"科学技术研究项目重点项目(吉教科合字[2016]179号);吉林省科技厅中青年领军人才及优秀创新团队项目(20111815)

摘　　要：生育酚环化酶(tocopherol cyclase,TC)是植物Vit E生物合成途径的关键酶之一。该研究根据红花转录组数据,利用RT-PCR(reverse transcription-PCR)方法从药用植物红花种子中克隆得到生育酚环化酶基因序列,命名为Ct TC,对其进行生物信息学分析,结果表明该基因开放阅读框(ORF)为1 524 bp,编码507个氨基酸,相对分子质量为62.9 k Da,等电点(theoretical p I)为5.01。跨膜结构分析表明TC存在跨膜结构。蛋白糖基化位点分析表明TC蛋白含有1个丝氨酸糖基化位点以及1个苏氨酸糖基化位点。亚细胞定位表明其定位于叶绿体中。利用荧光定量PCR检测红花不同发育时期种子以及干旱胁迫不同时间的红花叶片中TC基因的表达量,结果表明TC基因在红花的开花后50 d的种子中表达量最高,干旱胁迫6 d时TC基因的表达量达到最高值,随后下降,为TC基因在红花Vit E合成以及抵抗逆境胁迫中的功能研究奠定基础。The tocopherol cyclase was one of the key enzymes in plant vitamin E biosynthesis pathway. According to the study of Carthamus tinctorius transcriptome data,the Tocopherol cyclase gene was obtained using RT-PCR techniques and named Ct TC. Bioinformatics analysis showed theopen reading frame（ ORF） of Ct TC was 1 524 bp. The putative protein contained 507 amino acids with a predicted molecular mass of 62. 9 k Da and theoretically isoelectric point was 5. 01. Signal peptide analysis showed that it was a non secretory protein,and there was no signal peptide. The subcellular localization showed that the Ct TC protein was located in the chloroplast. The expression of Ct TC gene in safflower seeds at different development stages was determined by quantitative real-time PCR,it was found that the highest expression level of Ct TC gene was detected in 50 DAF. Quantitative RT-PCR analysis suggested that expression of Ct TC is induced and strengthened by drought stresses. This research provided a candidate gene for metabolic engineering of vitamin E and resisting stress.

关 键 词：红花 生育酚环化酶 克隆 表达分析 

分 类 号：S567.2[农业科学—中草药栽培]