The cloning of mouse FasL gene and expression in chondrocytes

AIM: To clone mFasL cDNA, then insert it into retrovirus expression system pLNCX 2, and express mFasL cDNA in chondrocytes to study its function. METHODS: RT PCR was applied to amplify mFasL cDNA from the total RNA of activated mouse spleen lymphocytes. The cDNA was inserted into a T cloning vector, then subcloned into pLNCX 2. After transfection of chondrocytes, expression of FasL cDNA was detected by FACS, and activity of expressed product was analyzed by single mixed lymphocyte reaction(SMLC). RESULTS: mFasL cDNA fragment of 0.855 kb was successfully cloned and verifiedby sequencing. The transfection efficiency of mFasL cDNA in chondrocytes determined by FACS was 60.64%. Expressed product could evidently inhibit SMLC of allogeneic lymphocytes. It’s stimulating index (SI) reduced to 11.71% as compare with the untransfected chondrocytes. CONCLUSION: Cloned recombinant pLNCX 2 FasL can effectively express mFasL in chondrocytes, and can evidently inhibit SMLC of allogeneic lymphocytes.