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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EcoR I)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。
Objective To construct eukaryotic expression vector containing mouse PD-L1 gene and obtain CHO cell line stably expressing mouse PD-L1 by transfection. Methods The PD-L1 gene was amplified from cDNA reverse transcribed from total RNA of mouse spleen cells and inserted into eukaryotic expression vector pIRES2-EGFP by double enzyme digestion (Xho I and EcoR I). CHO cells were transfected by lipofectamine After screening by G418, a CHO cell line stably expressing PD-L1 molecule was established. Results The eukaryotic expression vector pIRES2-EGFP / PD-L1 was constructed and a CHO cell line stably expressing the PD-L1 gene was established. Conclusion The successful construction of PD-L1 eukaryotic expression vector and the obtain of CHO cell line stably expressing the molecule laid the material foundation for subsequent research.