芥蓝八氢番茄红素脱氢酶基因BaPDS1的克隆及原核表达

发布时间：2018-02-03 17:07

  本文关键词： 芥蓝 八氢番茄红素脱氢酶 基因克隆 原核表达　出处：《基因组学与应用生物学》2016年07期 　论文类型：期刊论文

【摘要】：本研究以‘明丰香菇’芥蓝叶片总RNA为模板,采用同源克隆法设计并合成特异性引物,采用反转录PCR技术克隆得到芥蓝类胡萝卜素生物合成关键结构基因-八氢番茄红素脱氢酶基因Ba PDS1。序列分析表明,该基因c DNA序列长1 692 bp,编码563个氨基酸,推测蛋白等电点为5.96,理论分子量为63.15 k D;序列比对发现芥蓝Ba PDS1基因编码的氨基酸序列与甘蓝、花椰菜、油菜等植物的PDS具有很高的同源性;系统进化树分析表明,芥蓝PDS1与甘蓝PDS1蛋白的亲缘关系最为接近。之后构建p EASY-Blunt E1-Ba PDS1原核表达载体,转入Transetta(DE3)感受态细胞中,诱导后获得特异表达蛋白条带,且大部分以包涵体形式存在。本研究为揭示芥蓝Ba PDS1基因功能提供了一定的理论依据。
[Abstract]:In this study, the specific primers were designed and synthesized by homologous cloning using the total RNA of 'Mingfeng Lentinus edodes' leaves as template. The key structural gene of carotenoid biosynthesis of kale was cloned by reverse transcription PCR. The gene Ba PDS1 of lycopene dehydrogenase of octahydrolycopene was sequenced. The c DNA sequence is 1 692bpand encodes 563 amino acids. The deduced isoelectric point of the protein is 5.96 and the theoretical molecular weight is 63.15kD; Sequence alignment showed that the amino acid sequence encoded by Ba PDS1 gene of kale had high homology with PDS of cabbage, cauliflower, rape and so on. Phylogenetic tree analysis showed that the relationship between kale PDS1 and cabbage PDS1 protein was the most close, and then the prokaryotic expression vector of p EASY-Blunt E1-Ba PDS1 was constructed. Transfered into transettade3) cells, the specific expression protein bands were obtained after induction. Most of them exist in the form of inclusion bodies. This study provides a theoretical basis for revealing the function of Ba PDS1 gene in kale.
【作者单位】： 四川农业大学园艺学院;
【基金】：国家自然科学基金项目(31500247) 四川省教育厅重点项目(14ZA0016) 四川农业大学本科生科研兴趣培养计划项目(2015009)共同资助
【分类号】：S635.9
【正文快照】： Cloning and Prokaryotic Expression of Ba PDS1 Gene in Brassica alboglabraXue Shengling Zhang Fen Xia Xue Leng Zhenmin Chen Qing Tang Haoru Sun Bo*College of Horticulture,Sichuan Agricultural University,Chengdu,611130*Corresponding author,14099@sicau.edu.，

本文编号：1487906