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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
引用本文:付文哲,陈永井,王勤,张雪琨,马钰,张学光. 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[J]. 苏州大学学报(自然科学版), 2012, 0(2): 224-227
作者姓名:付文哲  陈永井  王勤  张雪琨  马钰  张学光
作者单位:苏州大学生物技术研究所,江苏苏州215007
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(XhoI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pIRES2-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。

关 键 词:PD-L1  真核表达载体  转染  CHO细胞

Construction of eukaryotic expressing vector of mouse PD-L1 and establishment of stable transfectant CHO cell line
FU Weng-zhe,CHEN Yong-jing,WANG Qin,ZHANG Xue-kun,MA Yu,ZHANG Xue-guang. Construction of eukaryotic expressing vector of mouse PD-L1 and establishment of stable transfectant CHO cell line[J]. Suzhou University Journal of Medical Science, 2012, 0(2): 224-227
Authors:FU Weng-zhe  CHEN Yong-jing  WANG Qin  ZHANG Xue-kun  MA Yu  ZHANG Xue-guang
Affiliation:(Biotechnology Institute of Soochow University, Jiangsu Suzhou 215007 ,China)
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 frag- ment 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-LI was transfected into CHO cells by Lipofectamine 2 000. Results The eukaryotic expressing vector pIRES2-EGFP/PD-L1 was constructed. Stable transfected CHO cell line expressing pD-L1 gene was established. Conclusion The construction of eukaryotic expressing vector pIRES2-EGFP/PD-L1 and the establishment of stable trans- fected CHO cell line have provided solid experimental foundation for further studies.
Keywords:PD-L1  eukaryotic expressing vector  transfection  CHO cell line
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