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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 34, No. 1, 2013, pp. 33-38
Bioline Code: zr13006
Full paper language: English
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 34, No. 1, 2013, pp. 33-38

 en Yeast-based production, purification and bioactivity assay of rainbow trout LECT2
SHI, Yu-Hong; ZHANG, Rui-Cheng; YANG, Dan-Yang; LU, Xin-Jiang; CHEN, Jiong & LI, Min-Yun

Abstract

Leukocyte cell-derived chemotaxin 2 (LECT2) is a secretory cytokine that functions in many physiological and pathological processes. We used a Pichia pastoris check for this species in other resources expression system for the recombinant expression of rainbow trout LECT2. The recombinant LECT2 was purified by UNOsphere S Cation exchange and size-exclusion chromatography columns. The obtained target protein was highly pure (>96% homogeneity) and the yield was >120 mg/L of yeast cultures. An in vitro chamber assay revealed that recombinant LECT2 could induce chemotactic responses in rainbow trout head kidney-derived macrophages. Recombinant LECT2 not only enhanced macrophage respiratory burst activity and bactericidal activity, but also changed macrophage gene expression. In summary, we established a rapid and efficient method to prepare active recombinant rainbow trout LECT2 using a yeast expression system and column chromatography. Bioactive recombinant LECT2 is essential for studies on protein functions.

Keywords
LECT2; Pichia pastoris; Purification; Bioactivity; Oncorhynchus mykiss

 
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