Abstract To construct, express and purify fusion protein containing HBcAg and HBsAg preS1 epitope peptide for the purpose of investigating a novel HBV vaccine with both prophylactic and therapeutic functions. Using DNA recombinant technology, we constructed prokaryotic expression plasmid pBTcs1 expressing HBcAg and HBsAg pre-S1 epitope peptide fusion protein. After expressed in E.coli. HB101, the production BTcs1 was purified by sucrose density gradient ultracentrifugation and identified by SDS-PAGE, SEC, WESTERN-BLOT and electron microscope. The results indicated that expression plasmid pBTcs1 was constructed successfully, and 20-25 mg purified BTcs1 fusion protein was obtained from 1 L LB culture. Result of DOT-BLOT indicated that the distribution of BTcs1 was mainly in 30-50% sucrose, the purity of BTcs1 was greater than 95% by SDS-PAGE and SEC analysis. BTcs1 could probe with specific antibodies at 28 kD by WESTERN-BLOT, BTcs1 could also self assemble VLP by electron microscope analysis, its diameter was 30-34 nm approximately. The present study lay a foundation for further research functions and applications of BTcs1.
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