[Construction and expression of eukaryotic expression plasmid pcDNA3.1/hIL-18].
OBJECTIVE
To construct an eukaryotic expression plasmid pcDNA3.1/hIL-18 and express it in mammalian cells.


METHODS
cDNA encoding mature hIL-18 was cleavaged by enzyme digestion from mesomeric clone vector pGEM-T/hIL-18 and inserted into an eukaryotic expression plasmid pcDNA3.1 to construct a recombinant expression plasmid pcDNA3.1/hIL-18. Then the constructed plasmid was transfected into COS-7 and Rlc310 cells by liposome-mediated gene transfer method. hIL-18 expressed in transfected COS-7 and Rlc310 cells was detected by immunohistochemical staining and level of hIL-18 mRNA in transfected Rlc310 cells was assayed by RT-PCR.


RESULTS
A recombinant eukaryotic expression plasmid pcDNA3.1/hIL-18 was successfully constructed and expressed transiently in COS-7 cells and stably in Rlc310 cells.


CONCLUSIONS
The construction and expression of pcDNA3.1/hIL-18 have been achieved successfully, which lays a foundation for further research on anti-tumor effect of IL-18.
Kong
Ling-Hong
LH
Institute for Cancer Research, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710061,China.
Li
Yao-Chen
YC
Wang
Yi-Li
YL
Shang
Ning-Kuan
NK
Lai
Bao-Chang
BC
Geng
Yi-Ping
YP
Si
Lu-Sheng
LS
D000818:Animals; D019556:COS Cells; D056890:Eukaryota / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D014162:Transfection