[1]薛生玲,江敏,常嘉琪,等.芥蓝牻牛儿基牻牛儿基焦磷酸合成酶基因BoaGGPPS1的克隆及表达分析[J].江苏农业学报,2018,(02):259-265.[doi:doi:10.3969/j.issn.1000-4440.2018.02.004]
　XUE Sheng-ling,JIANG Min,CHANG Jia-qi,et al.Cloning and expression analysis of geranylgeranyl pyrophosphate synthase gene（BoaGGPPS1） in Brassica oleracea var. alboglabra[J].,2018,(02):259-265.[doi:doi:10.3969/j.issn.1000-4440.2018.02.004]
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芥蓝牻牛儿基牻牛儿基焦磷酸合成酶基因BoaGGPPS1的克隆及表达分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年02期
页码:
259-265
栏目:
遗传育种·生理生化
出版日期:
2018-04-25

文章信息/Info

Title:
Cloning and expression analysis of geranylgeranyl pyrophosphate synthase gene（BoaGGPPS1） in Brassica oleracea var. alboglabra
作者:
薛生玲1; 江敏1; 常嘉琪2; 段妍雯1; 陈涛1; 郝宇1; 张芬1; 孙勃1
（1.四川农业大学园艺学院，四川成都611130；2.浙江大学农业与生物技术学院园艺系，浙江杭州310058）
Author(s):
XUE Sheng-ling1; JIANG Min1; CHANG Jia-qi2; DUAN Yan-wen1; CHEN Tao1; HAO Yu1; ZHANG Fen1; SUN Bo1
（1.College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China;2.Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China）
关键词:
芥蓝; 牻牛儿基牻牛儿基焦磷酸合成酶; 基因克隆; 表达分析
Keywords:
Chinese kale (Brassica oleracea var. alboglabra); geranylgeranyl pyrophosphate synthase; gene cloning; expression analysis
分类号:
S637.2;Q943.2
DOI:
doi:10.3969/j.issn.1000-4440.2018.02.004
文献标志码:
A
摘要:
本试验以芥蓝为试验材料，通过同源序列检索分析设计特异性引物，采用反转录PCR方法成功分离出芥蓝GGPPS1基因。BoaGGPPS1基因CDS全长为1 113 bp，编码370个氨基酸。生物信息学分析结果显示，BoaGGPPS1蛋白的等电点为6.07，相对分子量为39 790，不含跨膜结构域和信号肽。序列分析结果显示，BoaGGPPS1与其他植物GGPPS高度保守，与结球甘蓝的一致性最高，达到98%。系统进化树分析结果表明，BoaGGPPS1与十字花科其他植物的亲缘关系较近，与结球甘蓝的亲缘关系最近。BoaGGPPS1基因在芥蓝不同器官中均有表达，其中叶片中的表达量最高，根系中的表达量最低。构建原核表达载体pEASY-Blunt E1-BoaGGPPS1，并对其进行诱导表达，结果发现该蛋白在大肠杆菌体内主要以包涵体的形式存在。
Abstract:
Using Chinese kale (Brassica oleracea var. alboglabra) as experimental material, the BoaGGPPS1 gene encoding geranylgeranyl pyrophosphate synthase (GGPPS1) was successfully isolated by reverse transcription PCR and the specific primers were designed by homologous sequence analysis . The coding sequence length of BoaGGPPS1 gene was 1 113 bp, encoding 370 amino acids. Bioinformatics analysis results showed that the isoelectric point of BoaGGPPS1 protein was 6.07, and the relative molecular weight was 39 790, BoaGGPPS1 protein contained no transmembrane domain and signal peptide. The sequence analysis results showed that BoaGGPPS1 was highly conserved and had high identity with cabbage, reached 98%. Phylogenetic tree analysis results showed that BoaGGPPS1 was more closely related to other plants of Brassicaceae, and had the closest relationship with Brassica oleracea var. capitata. The expression of BoaGGPPS1 gene was found among different organs of Chinese kale, and the highest expression level was found in leaves，whereas the lowest expression was found in roots. The prokaryotic expression vector pEASY-Blunt E1-BoaGGPPS1 was constructed, and the protein was expressed in the form of inclusion bodies in Escherichia coli.

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备注/Memo

备注/Memo:
收稿日期：2017-09-15 基金项目：国家自然科学基金（31500247）；国家级大学生创新训练计划项目（201710626030） 作者简介：薛生玲（1993-），女，甘肃兰州人，硕士研究生，研究方向为蔬菜生理与分子生物学研究。（Tel）028-86291840；（E-mail）1181303872@qq.com 通讯作者：孙勃，（E-mail）14099@sicau.edu.cn
更新日期/Last Update: 2018-05-04