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[Construction and expression of human alpha-fetoprotein recombinant plasmid]
Xiao-ling Deng1, Mei-ying Cai, Da-peng Wei
1Department of Immunology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China. dengxiaoling@163.com
Objective:
To construct AFP-expressing plasmid and study the expression in eukaryotic cells.
Methods:
Total RNA was isolated from fetal liver tissue. Full-length human AFP cDNA was obtained by RT-PCR amplification and then recombined into eukaryotic expression plasmid pcDNA3.1. The AFP sequence of the recombinant plasmid phAFP was determined. The AFP expressions in CHO transfected with phAFP and in muscle after injection phAFP were investigated by immunohistological methods.
Results:
The restriction endonuclease mapping of recombinant plasmid showed 1.8 kb fragment as well as full-length human AFP cDNA, and the sequence is consistent with that from GenBank. The phAFP was successfully expressed in CHO and in muscle tissues.
Conclusion:
These results suggested that AFP can be used as a target gene of DNA vaccine in heptocellular carcinoma therapy.