Screening and Identification of ssDNA Aptamer for Human GP73

Figure 1

Preparation of the recombinant protein GP73. (a) The recombinant vector pET-32a-GP73 was confirmed by enzyme digestion. Lane 1, the double enzyme digestion product of the recombinant vector; Lane 2, the recombinant vector without double enzyme digestion. (b) The recombinant protein GP73 was induced in BL21 (DE3) by IPTG (1 mM) at 25°C. Lane 1, the supernatant of host BL21 (DE3) bearing pET-32a-GP73 with induction; Lane 2, the sediment of host BL21 (DE3) bearing pET-32a-GP73 with induction; Lane 3, the total lysate of BL21 (DE3) bearing pET-32a-GP73 without induction. (c) The recombinant protein GP73 was purified by affinity chromatography. Lane 1, the sample containing recombinant protein GP73 before loading affinity chromatography column; Lane 2, the eluate after loading affinity chromatography column; Lane 3, the eluate with PBS containing 20 mM imidazole; Lane 4, the eluate with PBS containing 40 mM imidazole; Lane 5, the eluate with PBS containing 60 mM imidazole; Lane 6, the eluate with PBS containing 80 mM imidazole; Lane 7, the eluate with PBS containing 100 mM imidazole; Lane 8, the eluate with PBS containing 500 mM imidazole. (d) The immunogenicity of the recombinant protein GP73 was confirmed by Western blotting with specific anti-GP73 antibody. Lane 1, the negative control; Lane 2, the purified GP73.

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