Construction expression and activity evaluation of a novel fusion toxin hIL15M/TNF?M
- VernacularTitle:新型重组毒素hIL15M/TNF?M的构建、表达与活性测定
- Author:
Fang LIU
;
Lei DING
;
Xiaohua MAO
;
- Publication Type:Journal Article
- Keywords:
human interleukin 15 mutant;
tumor necrosis factor alpha mutant;
protein fusion;
expression
- From:Journal of Third Military Medical University
2003;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct and express fusion toxin human IL15 mutant/TNF? mutant (hIL15M/TNF?M) in order to develop a targeted toxin capable of eliminating abnormal cells expressing high levels of hIL15 receptors. Methods DNA fragments of truncated hIL15M and TNF?M coding gene were joined in frame and cloned into T7 expression vector pET16b. The expressed fusion protein was purified from E. coli . Cytotoxicity of hIL15M/TNF?M was measured by MTT assay. Results E. coli transformed with the recombinant plasmid expressed an extra band with the expected size of 30?10 3. MTT assay indicated that PHA activated T cells were 13 times more sensitive to the cytotoxic effect of hIL15M/TNF?M than to the resting T cells. Conclusion hIL15M/TNF?M has high toxic effect against cells expressing high levels of IL15 receptors, suggesting that hIL15M/TNF?M has potential therapeutic effect on T cell leukemia and other diseases related with improperly activated T cells.
