[Construction and expression of human alpha-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.


CONCLUSIONS
These results suggested that AFP can be used as a target gene of DNA vaccine in heptocellular carcinoma therapy.
Deng
Xiao-ling
XL
Department of Immunology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China. dengxiaoling@163.com
Cai
Mei-ying
MY
Wei
Da-peng
DP
Hu
Li-juan
LJ
Bi
Jian-hong
JH
Yuan
Tun
T
D019496:Cancer Vaccines / Q000276:immunology / Q000627:therapeutic use; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D008113:Liver Neoplasms / Q000628:therapy; D010957:Plasmids / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection; D019444:Vaccines, DNA / Q000096:biosynthesis / Q000276:immunology; D000509:alpha-Fetoproteins / Q000096:biosynthesis* / Q000235:genetics*
D019496:Cancer Vaccines; D018076:DNA, Complementary; D011994:Recombinant Proteins; D019444:Vaccines, DNA; D000509:alpha-Fetoproteins