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Updated: Aug 20, 2026
Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5
Published on: January 28, 2016
[Cloning of mouse FasL full-length cDNA and construction of its recombinant adenovirus vector]
1Department of Clinical Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai 200433, China. yjhsmmu@yahoo.com.cn
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.
