[The cloning of mouse FasL gene and expression in chondrocytes].
OBJECTIVE
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 verified by 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.


CONCLUSIONS
Cloned recombinant pLNCX(2)-FasL can effectively express mFasL in chondrocytes, and can evidently inhibit SMLC of allogeneic lymphocytes.
Hu
Hong-Liang
HL
Key Laboratory of Tissue Engineering, Department of Plastic & Reconstructive Surgery, Affiliated Ninth People Hospital, Shanghai Second Medical University, Shanghai 200011, China.
Cao
Yi-Lin
YL
Liu
Yang
Y
Liu
Wei
W
Cui
Lei
L
Shang
Qing-Xin
QX
D000818:Animals; D019902:Chondrocytes / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D051379:Mice; D014162:Transfection
D018076:DNA, Complementary