烟草SKP1基因的原核表达  

Prokaryotic Expression for SKP1 Gene of Tobacco

作　　者：张付云[1,2] 赵小明[1] 白雪芳[1] 杜昱光[1] 

机构地区：[1]中国科学院大连化学物理研究所,辽宁省碳水化合物研究重点实验室,辽宁大连116023 [2]大连水产学院,辽宁大连116023

出　　处：《西北植物学报》2009年第8期1544-1549,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家“863”计划项目(2006AA10A213,2007AA091601);中国科学院知识创新工程重要方向项目(KSCX2-YW-G-041,KSCX2-YW-N-007);辽宁省自然科学基金项目(20082153)

摘　　要：为获取大量高纯度的枯斑三生烟SKP1蛋白以便研究其功能和性质,以pMD18-SKP1质粒为模板,PCR扩增枯斑三生烟的SKP1基因的cDNA编码区,经酶切后构建表达载体pET23b-SKP1,转入大肠杆菌BL21(DE3)进行诱导表达,并考察不同的IPTG诱导终浓度、表达时间和表达温度对目的蛋白His-SKP1表达量的影响。SDS-PAGE分析结果表明:最佳表达条件为不加诱导剂的情况下,37℃表达8 h。To obtain the SKP1 protein of tobacco with high purity in a large scale,the eDNA of SKP1 gene was amplified by PCR using pMD18-SKP1 as template,and was inserted into expression vector pET23b after being digested by restriction endonuclease. The recombinant plasmid pET23b-SKP1 was transformed into E. coli BL21 （DE3） for expression. In order to improve expression level of His-SKP1 ,the effects of various conditions were studied,including IPTG concentration, expression time and temperature. The results of SDS-PAGE analysis indicated that an ideally expression level could be obtained when the His-SKP1 were expressed for 8 hours at 37℃ without IPTG.

关 键 词：烟草 SKP1基因 原核表达 

分 类 号：Q789[生物学—分子生物学]