[Hepatocellular carcinoma associated antigen HCA520, a novel Ca2+-binding protein].
ABSTRACT To analyze the binding activity of the hepatocellular carcinoma associated antigen HCA520 to Ca2+.
The HCA520 gene was gained by PCR. Then, it was cloned into the prokaryotic expression vector pGEX-4T-3. The GST-HCA520 fusion gene was induced to express in E. coli and the expressed product was purified via GST-agarose affinity resin. The fusion protein was confirmed by Western blot analysis. The Ca2+ binding capability of the fusion protein was analyzed by dot blot.
The GST-HCA520 fusion gene was constructed and the protein was successfully expressed and purified, which was identified by DNA sequencing and Western blot respectively. The fusion protein could bind to Ca2+ depend on dosages.
HCA520, a novel hepatocellular carcinoma associated antigen, is a novel Ca2+ binding protein.