Cloning and expression of the extracellular domain of 4-1BBL.
- Author:
Wen-Guo JIANG
1
;
Dong-Sheng XIONG
;
Xiao-Feng SHAO
;
Jin-Hong WANG
;
Yuan-Fu XU
;
Fang LIU
;
Hong-Xing GUO
;
Zhen-Ping ZHU
;
Chun-Zheng YANG
Author Information
1. State Key Laboratory of Experimental Hematology , Institute of Hematology, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300020, China.
- Publication Type:Journal Article
- MeSH:
4-1BB Ligand;
biosynthesis;
genetics;
Apoptosis;
genetics;
Cell Line;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Extracellular Space;
metabolism;
Humans;
Interleukin-2;
biosynthesis;
Jurkat Cells;
Recombinant Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(5):703-707
- CountryChina
- Language:Chinese
-
Abstract:
RT-PCR was used to clone DNA fragment of the extracellular domain of 4-1BBL from human THP-1 cells (human monocyte), and the expression vector pAYZ4-1BBL was constructed by cloning the extracellular domain of 4-1BBL into the expression vector pAYZ. The extracellular domain of 4-1BBL was expressed in E. coli 16C9 and purified by affinity chromatography. SDS-PAGE and Western blot analysis showed that the relativae molecular weight of soluble 4-1BBL is 22kD which was consistent with the theoretically predicted value. So far as we know, it is the first time that the soluble expression of 4-1BBL in E. coli was achieved 4-1BBL induced a significant release of IL-2 in stimulated Jurkat cells after 48h incubation, especially in the presence of tumor cell. At the same time the apoptosis level of Jurkat cell reduce more than 50%. In conclusion, 4-1BBL may be useful in cancer immunotherapy.
