红花生育酚环化酶基因的克隆及表达分析

发布时间：2018-07-11 12:11

本文选题：红花 + 生育酚环化酶　；参考：《中国中药杂志》2016年20期

【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。
[Abstract]:Tocopherol cyclase (TC) is one of the key enzymes in plant VitE biosynthesis pathway. Based on the transcriptome data of safflower, the gene sequence of tocopherol cyclase was cloned from the seeds of safflower by reverse transcription-PCR. The results showed that the open reading frame (ORF) was 1 524 BP, encoding 507 amino acids, the relative molecular weight was 62.9 K Daand the isoelectric point (theoretical p I) was 5.01. The analysis of transmembrane structure shows that TC has transmembrane structure. The results of glycosylation site analysis showed that TC protein contained one serine glycosylation site and one threonine glycosylation site. Subcellular localization indicated that it was located in chloroplasts. Fluorescence quantitative PCR was used to detect the expression of TC gene in the seeds of different development stages of safflower and the leaves of safflower under different drought stress. The results showed that the expression of TC gene was the highest in the seeds 50 days after flowering of safflower. The expression of TC gene reached the highest level at the 6th day of drought stress, and then decreased, which laid a foundation for the study of the function of TC gene in the synthesis of vit E and resistance to stress in safflower.
【作者单位】：吉林农业大学生命科学学院;东北师范大学附属实验中学净月实验学校;
【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)
【分类号】：S567.2

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