Expression of macrophage activator Polytuftsin in Pichia pastoris and analysis of its biological activity

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Author:
MIAO Qing(State Key Laboratory of Molecular Oncology Cancer Institute Peking Union Medical College Chinese Academy of Medical Sciences, Beijing 100021, China)
YUAN Wei(State Key Laboratory of Molecular Oncology Cancer Institute Peking Union Medical College Chinese Academy of Medical Sciences, Beijing 100021, China)
WAN Yun-xia(State Key Laboratory of Molecular Oncology Cancer Institute Peking Union Medical College Chinese Academy of Medical Sciences, Beijing 100021, China)
MA Jie(State Key Laboratory of Molecular Oncology Cancer Institute Peking Union Medical College Chinese Academy of Medical Sciences, Beijing 100021, China)
Journal Title:
JOURNAL OF LEUKEMIA & LYMPHOMA
Issue:
Volume 18, Issue 05, 2009
DOI:
10.3760/cma.j.issn.1009-9921.2009.05.001
Key Word:
Macrophage-activating factors; Pichia; Leukemia

Abstract: Objective To investigate the secrection of Polytuftsin, a macrophage activator in Pichia pastoris expression system. Methods The recombinant plasmid encoding polytuftsin was transformed into the yeast by chemical method, and the positive clones were screened and expressed under the induction of methanod. It was observed that the influence of polytuftsin on macrophage in vivo and in vitro. Results It was verified that the recombinant products was confirmed to be our target protein by SDS-PAGE analysis and PCR assay. The expressed product could activate macrophage to secret cytokine and enhance the capacity of macrophages to kill L1210 tumor cells, the maximum killing rate is up to 40 %. In vivo the leukemia therapeutic trial, the time of tumor arise was delayed; the survival time of L1210 tumor-bear mice treated with Polytuftsin was extended, which is about 20 day. Conclusion The recombination protein polytuftsin, which could activate macrophage and has anti-tumor activity.

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