PLSCR1真核表达载体的构建及细胞内定位分析  

Construction of the eukaryotic expression vector of PLSCR1 and analysis of its intracellular localization

作　　者：王远[1,2] 陈英[2] 张学清[2] 陈忠民[2] 田喜凤[1] 陈晶[1] 

机构地区：[1]河北联合大学,河北唐山063000 [2]北京军事医学科学院,北京100850

出　　处：《生物学杂志》2015年第3期1-3,8,共4页Journal of Biology

基　　金：国家自然科学基金青年基金项目(81402631)

摘　　要：分别构建带有myc标签和GFP荧光蛋白的磷脂爬行酶1(PLSCR1)真核表达载体,获得两个融合表达载体,并转入HEK293细胞观察表达情况及细胞内定位,为研究PLSCR1的定位与功能的关系奠定基础。以本实验室保存的Hela c DNA文库为模板,采用PCR技术扩增PLSCR1编码序列,将其分别插入p CMV-Myc-N和p EGFP-C1载体,Western blotting检测其在HEK293中的表达,采用激光共聚焦观察p EGFP-C1融合表达载体在HEK293细胞中定位。通过DNA序列分析,证实了成功构建了PLSCR1真核表达载体,并能在HEK293细胞中实现基因的过表达。成功构建PLSCR1真核表达载体,为进一步研究其功能奠定了基础。Two eukaryotic expression vectors of phospholipid scramblase 1（PLSCR1）with myc-tag or Green Fluorescent Protein （GFP）, were constructed to obtain two fusion expression vectors, which were transfected to HEK293 cell, to observe the expression and cellular localization. The results would lay a foundation for the study of PLSCR1 gene localization and functional relationships. Hela cDNA library preserved in our laboratory was used as template, the PLSCR1 coding sequence was amplified by PCR and respectively inserted into the vector pCMV-Myc-N and pEGFP-C1. The epression was detected in HEK293 by Western blotting and localization of pEGFP-C1 fusion expression vector in HEK293 cells by laser scanning confocal microscopy. As results, the eukaryotic expression vector of PLSCR1 was successfully constructed by the DNA sequence analysis, and over expressed genes in HEK293 cells. It makes good foundation for further study of functions by successfully constructing eukaryotic expression vector of PLSCR1.

关 键 词：磷脂爬行酶1 真核表达载体 HEK293细胞 

分 类 号：Q78[生物学—分子生物学]