[Yeast-based production, purification and bioactivity assay of rainbow trout LECT2].
Dongwuxue Yanjiu
; 34(1): 33-8, 2013 Feb.
Article
em Zh
| MEDLINE
| ID: mdl-23389976
ABSTRACT
Leukocyte cell-derived chemotaxin 2 (LECT2) is a secretory cytokine that functions in many physiological and pathological processes. We used a Pichia pastoris 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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Contexto em Saúde:
3_ND
Problema de saúde:
3_neglected_diseases
/
3_zoonosis
Assunto principal:
Proteínas Recombinantes
/
Oncorhynchus mykiss
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Proteínas de Peixes
/
Peptídeos e Proteínas de Sinalização Intercelular
Limite:
Animals
Idioma:
Zh
Revista:
Dongwuxue Yanjiu
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
China