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
in Chinese
| WPRIM
| ID: wpr-332388
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Recombinant Proteins
/
Von Willebrand Factor
/
Transfection
/
Gene Expression
/
Cricetulus
/
CHO Cells
/
Cloning, Molecular
/
Trinucleotide Repeats
/
Genetic Vectors
Type of study:
Prognostic study
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Journal of Experimental Hematology
Year:
2010
Type:
Article
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