[Construction of bi-cistronic co-expression plasmid of mIL-12 and the expression in vitro or in vivo]
OBJECTIVE: To construct a bi-cistronic co-expression plasmid for mouse interleukin-12 and to observe its expression in vitro or in vivo.METHODS: The full-length cDNA encoding p35 and p40 was cloned into eukaryotic cells expression vector pcDNA 3.1 respectively. Subsequently,the p35 expression unit was inserted into pcDNA 3.1/p40 to produce the bi-cistronic co-expression plasmid in which the p35 and p40 genes were controlled by their own CMV.The plasmid was expressed in vitro and in vivo. RESULTS: The mIL-12 in the supernatant was detected by ELISA after the pCmIL-12 was transfected into COS-7 cells. The activity of NK cells could be augmented by the supernatant in vitro and also by by intradermal delivery of pCmIL-12 in vivo. CONCLUSION: The plasmid constructed by us can express biologically active mIL-12 in vitro and in vivo.
Chen
Wei
W
College of Medical Sciences, Zhejiang University, Hangzhou 310031, China.
Shao
Chuan-Sen
CS
Shen
Jian-Gen
JG
Bao
Jian-Fang
JF
Pan
Jian-Ping
JP
Han
Wei
W
Shou
Lin
L
Xiang
Shao
S
Zheng
Shu
S