Expression of transthyretin gene in Pichia pastoris

Plasmid pSK-TTR was digested by BamHI and the DNA fragment containing Transthyretin gene was cloned into the BamHI site of vector pHIL-SI. The recombinant plasmid pHIL-SI-TTR was digested by BglII; the larger fragment was transformed into yeast Pichia pastoris. SDS-PAGE analysis showed that the yeast transformant can express and secrete TTR as a fusion protein, TTRF. To purify TTRF,the DEAE-Sepherose F.F. chromatography and Sephacryl S-200 chromatography were used. The in vitro test showed that TTRF could inhibit the growth of human hepatoma cells.