Abstract

Objective To clone and express osteopontin (OPN) fusion proteins with high immunogenicity.Methods OPN gene was analyzed using Lasergene software,and a high immunogenic region was selected as target fragment.The fragment of the target gene was cloned by RT-PCR from total RNA,The amplified gene was inserted into vectors pET-32a and pGEX-4T-1.The recombinant plasmids were constructed and expressed proteins (including His-Tag and GST-Tag,respectively) were identified by SDS-PAGE and Western blotting.Results A DNA band about 351 bp was confirmed as target OPN fragment by DNA sequencing.Soluble expression of high immunogenic OPN fusion proteins was achieved,which was identified by Western blotting.Conclusion High immunogenic OPN fusion proteins with high purity have been expressed successfully in prokaryotic system.The study lays a foundation for further development of OPN diagnostic reagent. Key words: Osteopontin; Hepatocellular carcinomas; High immunogenic region

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