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.
Ying
C
C
Institute of Genetics Fudan University, Shanghai, China.
Boyu
G
G
Daxin
S
S
Hanying
Y
Y
Yongqin
C
C
Yuyang
L
L
Xiang
L
L
Peikun
T
T
Jianren
G
G
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001483:Base Sequence; D001681:Biological Assay; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy; D004305:Dose-Response Relationship, Drug; D005822:Genetic Vectors; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics; D011228:Prealbumin / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D011228:Prealbumin; D011993:Recombinant Fusion Proteins