Fusion protein VT-GL-B3, coding gene thereof and applications thereof

[0037] Embodiment 1, the preparation of VT-GL-B3 fusion protein

[0038] 1. Construction of recombinant plasmids

[0039] Insert the double-stranded DNA molecule shown in Sequence 2 of the sequence table between the NheI and NotI restriction sites of the pIRES2-EGFP vector to obtain the recombinant plasmid pIRES2-EGFP-VT-GL-B3. The schematic diagram of the structure of the recombinant plasmid pIRES2-EGFP-VT-GL-B3 is shown in figure 1 .

[0040] The double-stranded DNA molecule shown in Sequence 2 of the Sequence Listing was named VT-GL-B3 gene. The double-stranded DNA molecule shown in sequence 2 of the sequence listing encodes the protein shown in sequence 1 of the sequence listing. The protein shown in Sequence 1 of the sequence listing is named VT-GL-B3 fusion protein.

[0041] In sequence 1 of the sequence listing, amino acid residues 1 to 20 from the N-terminal are signal peptides, and amino acid residues 21-236 are VEGFR1D2R2D3 fragments (VEGFR1D2R2D3 fragments consist

[0056] Example 2, the biological activity of VT-GL-B3 mature peptide

[0057] 1. Affinity between VT-GL-B3 mature peptide and rhVEGF165

[0058] Biacore T200, CM5 sensor chip (Lot No.10148134), Amine Coupling Kit (Lot No.2057704), Human Antibody Capture Kit (Lot No.10137214), HBS-EP+buffer (10×) (Lot No.BCBJ0266V) Both were purchased from GE Healthcare.

[0059] Anti-Human IgG(Fc) Antibody is a component of Human Antibody Capture Kit (Lot No.10137214). HBS-EP + Buffer (10×) is diluted to 10 times volume with deionized water, which is HBS-EP + buffer solution.

[0060] 1. Anti-Human IgG (Fc) Antibody combined with the surface of CM5 sensor chip

[0061] (1) Embed the CM5 sensor chip into Biacore T200.

[0062] (2) Prepare Anti-Human IgG(Fc) Antibody solution with pH 5.0, 10mM sodium acetate buffer.

[0063] (3) According to the sample placement method in the Immobilization program setting, place the solution in the Amine Coupling Kit in the appropriate position of the Biaco