[Cloning of mouse FasL full-length cDNA and construction of its recombinant adenovirus vector]

AIM: To clone mouse FasL full-length cDNA and construct recombinant adenovirus expression vector.

METHODS: Mouse FasL full-length cDNA was cloned from BALB/c mouse's splenic mononuclear cells stimulated with ConA (5 mg/L) for 48 h, and then the adenovirus shuttle vector mFasL-pAdTrack containing mouse FasL full-length cDNA under the control of CMV promoter was constructed. The shuttle vector was linearized with Pme I and cotransformed into E.coli BJ5183 with pAdEasy-1, the adenovirus background plasmid vector. The positive recombinants were subsequently identified by restriction enzyme digestion, and transfected into HEK293 cells for preparing recombinant FasL adenoviruses.

RESULTS: The mouse FasL full-length cDNA has been cloned and recombinant mFasL adenovirus has been constructed successfully, as shown by PCR, restriction endonuclease digestion analysis and fluorescence detection.

CONCLUSION: The mouse FasL recombinant adenovirus prepared in this study can be used to study the role of FasL in tumor and autoimmune diseases.