摘要
本研究以芥蓝为实验材料,采用同源克隆的方法分离出Ba DXS1基因,其开放阅读框为2 139 bp,编码712个氨基酸。理化性质分析表明,Ba DXS1蛋白的分子量为77.21 ku,等电点p I为8.59,不含跨膜区域,位于植物细胞叶绿体内。氨基酸序列比对发现,芥蓝DXS1与甘蓝、拟南芥、烟草、番茄等植物的DXS蛋白序列的一致性达到79%以上;系统进化树分析显示,芥蓝DXS1与甘蓝DXS的亲缘关系最近。构建原核表达载体p EASY-Blunt E1-Ba DXS1,并对其进行诱导表达,发现该蛋白在大肠埃希菌体内主要以包涵体的形式存在。
The 1-deoxy-D-xylulose-5-phosphate synthase（ DXS） encoding gene Ba DXS1 was isolated from Chinese kale（ Brassica alboglabra） leaves by using homologous cloning method. Ba DXS1 contained an open reading frame of2 139 bp in length,encoding 712 amino acids. Physical and chemical properties analysis indicated that the molecular weight of Ba DXS1 protein is 77. 21 ku,and the isoelectric point p I is 8. 59. There was no transmembrane region,and Ba DXS1 is located in chloroplast of plant cell. Amino acid sequences comparison results showed that the identities of Ba DXS1 and those in Brassica oleracea,Arabidopsis thaliana,tobacco,tomato were all over 79%. Phylogenetic tree analysis showed that it had close relation with Bo DXS. The prokaryotic expression vector p EASY-Blunt E1-Ba DXS1 was constructed,and the protein was successfully expressed in the form of inclusion bodies in Escherichia coli.
作者
薛生玲
江敏
常嘉琪
刘洋
魏淋
周建坤
张芬
孙勃
XUE Shengling1, JIANG Min1,CHANG Jiaqi2,LIU Yang 1,WEI Lin 1 ,ZHOU Jiankun1,ZHANG Fen1,SUN Bo1(1. College of Horticulture,Sichuan Agricultural University, Chengdu 611130, China; 2. Department of Horticul-ture, Zhejiang University,Hangzhou 310058,Chin)
出处
《浙江农业学报》
CSCD
北大核心
2018年第5期771-777,共7页
Acta Agriculturae Zhejiangensis
基金
国家自然科学基金(31500247)
四川省省级大学生创新训练计划项目(201710626030)
