Cloning and eukaryotic expression vector construction of mouse FasL gene and its expression in dendritic cells
- VernacularTitle:小鼠FasL全长cDNA的克隆、真核表达载体构建及其在树突状细胞中的表达
- Author:
Yan WANG
- Publication Type:Journal Article
- Keywords:
FasL;
clone;
vector construction;
dendritic cell
- From:Journal of Third Military Medical University
1984;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone mouse FasL(mFasL) full length cDNA and construct eukaryotic expression vector,and to detect its expression in dendritic cells.Methods RT-PCR and TA cloning technique were used to clone the mFasL full length cDNA from mouse activated T lymphocytes,and then eukaryotic expression vector pIRES2EGFP containing mFasL cDNA was constructed.The positive recombinants pIRES2EGFP-mFasL were transfected into dendritic cells(DC),the expression of mFasL mRNA was detected by RT-PCR and the expression of mFasL protein was assayed by immunofluorescence and Western blotting.Results The cloned full reading frame of mFasL cDNA was in coincidence with the sequence registered in GenBank.In the DC after transfection of pIRES2EGFP-mFasL,the expressions of mFasL mRNA and mFasL protein were detected.Conclusion The mFasL cDNA was cloned and constructed into the eukaryotic expression vector successfully,and mFasL gene could be expressed efficiently in DC transfected by pIRES2EGFP-mFasL.
