巴西橡胶树K+通道蛋白HbKCO1的原核表达

中国农学通报 ›› 2009, Vol. 25 ›› Issue (13): 33-36.

所属专题：生物技术；园艺

巴西橡胶树K+通道蛋白HbKCO1的原核表达

蔡元保,,朱家红,张全琪,,张治礼,

收稿日期:2009-04-28 修回日期:2009-05-07 出版日期:2009-07-05 发布日期:2009-07-05

Prokaryotic Expression of HbKCO1，a K+ channel protein from Hevea brasiliensis

Cai Yuanbao,, Zhu Jiahong, Zhang Quanqi,, Zhang Zhili,

Received:2009-04-28 Revised:2009-05-07 Online:2009-07-05 Published:2009-07-05

摘要/Abstract

摘要： K+通道蛋白在维持细胞离子平衡等生命活动中发挥重要的作用。利用真核或原核表达系统高效表达K+通道蛋白是深入研究其生化特征及生理功能的基础和前提。本研究在成功克隆巴西橡胶树(Hevea brasiliensis)K+通道蛋白基因cDNA全长的基础上，将HbKCO1 cDNA 编码区插入pET-28a构建原核表达载体pET-28a(+)-HbKCO1，并分别转化大肠杆菌RosettaTM(DE3) pLysS和BL21(DE3)菌株。经过条件优化，转化的大肠杆菌RosettaTM(DE3) pLysS菌株经1mM IPTG诱导3h能够高效表达HbKCO1重组蛋白，但同等条件下转化的BL21(DE3)菌株仅能微量表达。HbKCO1重组蛋白原核表达具有菌株依赖性。

关键词: 水培, 水培, 生菜, 营养液

Abstract: K+ channel proteins play key roles in ion balance in plant cells. With the help of eukaryotic or prokaryotic expression system, efficient expression of K+ channel protein is a prerequisite for further study of its biochemical characteristics and physiological functions. Here, a K+ channel gene，HbKCO1 was isolated from Hevea brasiliensis，and expression of recombinant HbKCO1 was investigated. ORF fragment of HbKCO1 cDNA was inserted into the vector pET-28a(+), resulting in pET-28a(+)-HbKCO1, and then pET-28a(+)-HbKCO1 was transformed in Escherichia coli RosettaTM(DE3) pLysS and BL21(DE3), respectively. In the optimum condition, recombinant HbKCO1 could be efficiently expressed in the E.coli RosettaTM(DE3) pLysS transformats after 3h induction of 1mM IPTG, but weakly expressed in the E.coli BL21(DE3) transformats. Prokaryotic expression of recombinant HbKCO1 was strain-dependent.

蔡元保,,朱家红,张全琪,,张治礼,. 巴西橡胶树K+通道蛋白HbKCO1的原核表达[J]. 中国农学通报, 2009, 25(13): 33-36.

Cai Yuanbao,, Zhu Jiahong, Zhang Quanqi,, Zhang Zhili,. Prokaryotic Expression of HbKCO1，a K+ channel protein from Hevea brasiliensis[J]. Chinese Agricultural Science Bulletin, 2009, 25(13): 33-36.

参考文献

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巴西橡胶树K+通道蛋白HbKCO1的原核表达

Prokaryotic Expression of HbKCO1，a K+ channel protein from Hevea brasiliensis

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