The invention relates to method for preparing
venom edema factor inhibition
protein and the application thereof, belonging to the field of
biotechnology and relating to a method for separating and purifying a
protein having inhibition action on
venom edema factors from the
blood serum of Gloydius shedaoensis. The invention is characterized by the method adopted for separating and purifying the inhibition
protein, and the method comprises the following steps: performing the
ammonium sulphate
precipitation on the crude
blood serum of Gloydius shedaoensis, performing gel
filtration twice, passing through an
anion exchange column, an
affinity chromatography column and a reversed phase HPLC to obtain a
single component with the molecular weight of 25KDa, and identifying 70
amino acid sequencesof the N terminal and 20
amino acid sequences of the C terminal of the protein. The invention is also characterized by obtaining the
gene sequence corresponding to the
venom edema factor inhibition protein, the
gene sequence is established on a TOPO
expression vector for producing
in vitro and expressing the protein. The invention has the advantages of providing a preparation method for producingin vitro venom edema factor inhibition protein, and the inhibition protein can be applied to the
clinical treatment of snakebite and other diseases.