PC-1在前列腺癌细胞系LNCaP中的定位  

Subcellular Localization of PC-1 in LNCaP Prostate Cancer Cells

作　　者：伍飞马[1,2] 钱晓龙[1] 施庆国[1] 李山虎[1] 王洪涛[1] 祁建民[2] 周建光[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]福建农林大学生命科学院,福建福州350002

出　　处：《生物技术通讯》2012年第1期15-19,共5页Letters in Biotechnology

基　　金：国家自然科学基金(30770834;30870961);国家高技术研究发展计划(2008AA02Z123)

摘　　要：目的:研究PC-1分子在前列腺癌细胞系LNCaP中的亚细胞定位。方法:利用常规PCR和重叠PCR技术在pEGFP-C1-PC-1上分别扩增PC-1的不同截短体及缺失体基因,PCR产物经酶切后克隆到真核表达载体pEGFP-C1中;经测序确定构建成功的载体在LNCaP细胞中瞬时高表达,在荧光显微镜下观察这些载体表达产物在LNCaP细胞中的定位情况,并通过Western印迹验证这些载体在LNCaP细胞中的表达。结果:构建了多个用于PC-1亚细胞定位研究的、能在LNCaP细胞中表达的载体;同时,还找到了一段对PC-1定位有重要影响的氨基酸序列。结论:为进一步研究PC-1的亚细胞定位及其发挥功能的方式提供了基础。Objective： To study the subcellular localization of PC-1 in LNCaP prostate cancer cells.Methods： A various of PC-1 segments were amplified and cloned into a eukaryotic expression vector pEGFP-C1,respectively.The sequencing recombinants were transfected into LNCaP cells,observed the distribution of green fluorescence in the cell and validated their expressions by Western blot.Results： The recombinant vectors used for subcellular localization study of PC-1 had been successfully constructed and expressed in LNCaP cell lines.Meanwhile,a sequence that significantly impacts the subcellular localization of PC-1 had been identified.Conclusion： This work provides the basis for further study of subcellular localization and way of functioning of PC-1.

关 键 词：PC-1 亚细胞定位 LNCaP细胞载体构建 重叠PCR 基因表达 

分 类 号：Q78[生物学—分子生物学] Q23