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


CONCLUSIONS
The mouse FasL recombinant adenovirus prepared in this study can be used to study the role of FasL in tumor and autoimmune diseases.
Yang
Jia-Hui
JH
Department of Clinical Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai 200433, China. yjhsmmu@yahoo.com.cn
Shen
Qian
Q
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics* / Q000378:metabolism; D053222:Fas Ligand Protein / Q000235:genetics*; D005818:Genetic Engineering; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics
D018076:DNA, Complementary; D004274:DNA, Recombinant; D053222:Fas Ligand Protein; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D004262:DNA Restriction Enzymes