人程序性死亡配体2基因真核表达载体的构建及其在L929细胞株中的表达  

Construction of Eukaryotic Expressing Vector of Human PD-L2 and Its Expression in L929 Cell Line

作　　者：沈冬[1] 张瑜[2] 

机构地区：[1]苏州市中医医院检验科,江苏苏州215009 [2]苏州市立医院北区检验科,江苏苏州215008

出　　处：《中国血液流变学杂志》2015年第4期406-408,441,共4页Chinese Journal of Hemorheology

摘　　要：目的：构建人程序性死亡配体2（PD-L2）基因的真核表达载体，并通过转染获得稳定表达该分子的L929基因转染细胞株。方法获取人外周血单个核细胞，通过RT-PCR方法获取PD-L2基因全长序列。并与pIRES2真核表达载体连接，通过脂质体方法转染L929细胞，经G418筛选获得稳定表达该基因的细胞株。流式细胞术鉴定PD-L2基因的表达。结果成功构建了真核表达载体pIRES2/PD-L2，建立了稳定转染该基因的L929细胞株。结论稳定表达PD-L2分子的基因转染细胞株为进一步研究该分子的生物学功能提供了物质基础。Objective To construct eukaryotic expressing vector of program death ligand-2 （PD-L2） gene and transfect to L929 cell line so as to establish a cell line stably expressing PD-L2 gene.MethodsThe full-length human PD-L2 gene were cloned from human PBMC through RT-PCR methods and then inserted into the eukaryotic expression vector pIRES2-EGFP to construct the recombinant pIRES2-EGFP/PD-L2. Recombinant plasmid was transfected into murine L929 cells after induction with LipofectamineTM 2000 Reagent, followed by G418 selection.ResultsThe eukaryotic expressing vector pIRES2-EGFP/PD-L2 was constructed stable transfected L929 cell line was established and PD-L2 gene was expressed successfully.Conclusion The construction of eukaryotic expressing vector pIRES2-EGFP/PD-L2 and the development of stable transfected L929 cells have provided a solid experiment foundation for further studies of PD-L2 function.

关 键 词：PD-L2基因 真核表达载体 转染 L929细胞 PD-L2 

分 类 号：R730.3[医药卫生—肿瘤]