CAJ | 학술논문

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 cDNA文库为模板，采用PCR技术扩增PLSCR1编码序列，将其分别插入pCMV-Myc-N和pEGFP-C1载体， Western blotting检测其在HEK293中的表达，采用激光共聚焦观察pEGFP-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。
분별구건대유myc표첨화GFP형광단백적린지파행매1（PLSCR1）진핵표체재체，획득량개융합표체재체，병전입HEK293세포관찰표체정황급세포내정위，위연구PLSCR1적정위여공능적관계전정기출。이본실험실보존적Hela cDNA문고위모판，채용PCR기술확증PLSCR1편마서렬，장기분별삽입pCMV-Myc-N화pEGFP-C1재체， Western blotting검측기재HEK293중적표체，채용격광공취초관찰pEGFP-C1융합표체재체재HEK293세포중정위。통과DNA서렬분석，증실료성공구건료PLSCR1진핵표체재체，병능재HEK293세포중실현기인적과표체。성공구건PLSCR1진핵표체재체，위진일보연구기공능전정료기출。
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.