NtSKP1-GFP植物表达载体的构建及亚细胞定位  

Construction of NtSKP1-GFP Plant Expression Vector and Subcellular Location

作　　者：张付云[1,2] 陈士云[3] 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,大连116023 [2]大连水产学院,大连116023 [3]中国科学院武汉病毒研究所,武汉430071

出　　处：《西北农业学报》2009年第4期144-148,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家高技术研究发展计划"863"(2001AA620501);(2002AA245131)资助

摘　　要：用PCR技术扩增NtSKP1基因的编码区,定向克隆至表达载体pCAMBIA1302上构建用于瞬时表达NtSKP1蛋白的重组质粒。重组质粒经PCR和测序鉴定后用冻融法转入农杆菌LBA4404,进而经农杆菌LBA4404介导转入烟草悬浮细胞,激光共聚焦显微镜观察确定其亚细胞定位。测序结果表明,插入片段与预期序列完全一致,与载体形成了一个完整的基因表达盒;亚细胞定位结果表明NtSKP1蛋白在胞浆和核部位均有分布。To construction plant expression vector of NtSKP1, the open reading frame of NtSKP1 was amplified by PCR, and was inserted to plasmid pCAMBIA1302. DNA sequencing showed that the sequence of the recombinant plasmid was correct. Then the recombinant plasmid was transferred into Agrobacteriurn turnefaciens strain LBA4404 through freeze-thaw method. To subcellular location the NtSKP1, the recombinant plasmid was transformed into the tobacco suspension cells mediated by Agrobacterium tumefaciens strain LBA4404, and the location of NtSKP1 was detected in both cytoplasm and nucleus.

关 键 词：NtSKP1基因 植物表达载体 亚细胞定位 

分 类 号：S184[农业科学—农业基础科学]