[Co-expression of HSV-1 gD gene and interleukin-2 gene in dendritic cell].
OBJECTIVE
To construct eukaryotic expression plasmid IRES-gD-IL-2 which contains both HSV-1 glycoprotein D (gD) gene and IL-2 gene and to induce it to express in antigen presenting cell (APC) -- dendritic cells (DCs).


METHODS
The whole sequence of gD and IL-2 were amplified by PCR assay. After confirmation by PCR, double-enzyme digestion and sequencing, these genes were directly cloned into eukaryotic expression vector IRES, then were transfected into DCs. Western blotting was employed to identify the transcription and expression of gD gene.


RESULTS
The results of PCR and enzyme digestion showed that the recombinant expression plasmid contained correct fragments, and the transcription and expression of gD were confirmed by Western blotting.


CONCLUSIONS
The recombinant expression vector IRES-gD-IL-2 was constructed, the results of the Western blotting showed that the recombinant protein could be identified by gD- specific antibody, therefore the protein has immunologic competence.
Li
Wei-ong
WO
Department of Neurology, Beijing Friendship Hospital, Beijing 100050, China.
Wang
De-xin
DX
Wang
Jian-wei
JW
Niu
Bo
B
Xu
Xian-min
XM
D015153:Blotting, Western; D003713:Dendritic Cells / Q000378:metabolism*; D022301:Herpes Simplex Virus Vaccines / Q000276:immunology; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology; D019444:Vaccines, DNA / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics*
D022301:Herpes Simplex Virus Vaccines; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D014759:Viral Envelope Proteins; C515577:glycoprotein gD, herpes simplex virus type 1