[Construction of recombinant adenovirus expressing sCD40L-Ig].
OBJECTIVE
To construct adenovirus expressing secretory human CD40L-Ig (sCD40 ligand Ig fusion protein).


METHODS
The genes encoding the extracellular domain of human CD40L and IgG Fc were amplified with PCR and inserted into shuttle vector pAdTrack-CMV which then was transformed into E. coli pAdEasy-1-BJ5183 to produce recombinant Ad plasmid. The recombinant plasmid was digested with Pac I and transfected into 293 cells to generate recombinant adenovirus. The recombinant adenovirus was then used in mixed lymphocyte reaction (MLR) to test its function.


RESULTS
The recombinant adenovirus sCD40L-Ig was produced. It could inhibit the lymphocyte proliferation in MLR.


CONCLUSIONS
The sCD40L-Ig adenovirus is prepared successfully and its inhibition of MLR is confirmed.
Li
Zhao-lun
ZL
Renal Disease Center of First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China. Lizhaolun1@sina.com.cn
Tian
Pu-xun
PX
Xue
Wu-jun
WJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*
C526581:CD40LIg fusion protein; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins