小鼠PD-L1真核表达载体的构建及其在CHO细胞中的稳定表达
Construction of eukaryotic expressing vector of mouse PD-L1 and establishment of stable transfectant CHO cell line
付文哲 1陈永井 1王勤 1张雪琨 1马钰 1张学光1
作者信息
- 1. 苏州大学生物技术研究所,江苏苏州215007
- 折叠
摘要
目的 构建含有小鼠PD-L1基因的真核表达载体,并通过转染获得稳定表达小鼠PD-L1分子的CHO细胞系.方法 从小鼠脾细胞总RNA逆转录的cDNA中扩增出PD-L1基因,通过双酶切(Xho Ⅰ和EcoRⅠ)装入真核表达载体pIRES2-EGFP中,脂质体法转染CHO细胞,经G418筛选后,建立稳定高表达PD-L1分子的CHO细胞系.结果 构建了真核表达载体plRES2-EGFP/PD-L1,建立了稳定表达PD-L1目的基因的CHO细胞系.结论 成功构建了 PD-L1真核表达载体并获得了稳定表达该分子的CHO细胞系为后续研究奠定了物质基础.
Abstract
Objective To construct eukaryotic expressing vector of mouse PD-L1 gene and transfect CHO cells so as to establish stable CHO cell line expressing PD-L1. Methods Mouse PD-L1 gene fragment was obtained by RT-PCR, recombinant vector was constructed by linking the PCR products and pIRES2-EGFP which was digested by Xho I and EcoR I. Then pIRES2-EGFP/PD-Ll was transfected into CHO cells by Lipofectamine 2 000. Results The eukaryotic expressing vector pIRES2-EGFP/PD-Ll was constructed. Stable transfected CHO cell line expressing PD-L1 gene was established. Conclusion The construction of eukaryotic expressing vector pIRES2-EGFP/PD-Ll and the establishment of stable transfected CHO cell line have provided solid experimental foundation for further studies.
关键词
PD-L1/真核表达载体/转染/CHO细胞Key words
PD-L1/eukaryotic expressing vector/transfection/CHO cell line引用本文复制引用
出版年
2012
NETL
NSTL
维普
万方数据