Eukaryotic expression of von Willebrand factor A1A2A3 triplet and its biological activity / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 1224-1228, 2010.
Article
em Zh
| WPRIM
| ID: wpr-332388
Biblioteca responsável:
WPRO
ABSTRACT
The aim of this study was to construct the eukaryotic expression vectors harboring human vWF-A1A2A3 gene and to investigate its expression in CHO cells and biologic function so as to provide a basis for further exploring the biologic activity of vWF-A1A2A3. The primers were designed according to published sequences; the human vWF-A1A2A3 was amplified by PCR from vWF cDNA; the fragment of interest was inserted into eukaryotic expression vector pSectag2b by using restriction enzyme and ligase after vWF-A1A2A3 was confirmed by sequencing. The recombinant expression plasmid was transfected into CHO cells and the stable expression product (rvWF-A1A2A3) was detected by using Western blot. The results showed that the eukaryotic expression vector pSectag2b-A1A2A3 was successfully constructed and expressed its corresponding protein efficiently. The recombinant protein was identified to be able to bind collagen and ristocetin-induced platelets. It is concluded that the pSectag2b-A1A2A3 is successfully constructed and can express in CHO cells. The rvWF-A1A2A3 protein established in this study provides a basis for the further study on its biological structure, function and clinical application.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes
/
Fator de von Willebrand
/
Transfecção
/
Expressão Gênica
/
Cricetulus
/
Células CHO
/
Clonagem Molecular
/
Repetições de Trinucleotídeos
/
Vetores Genéticos
Tipo de estudo:
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Journal of Experimental Hematology
Ano de publicação:
2010
Tipo de documento:
Article