Construction and Expression of Human α-Fetoprotein Recombinant Plasmid

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.