Expression and purification of a putative tumor suppressor gene PP5715 in E. coli with growth inhibition of hepatocellular carcinoma cells.
PP5715 is a putative tumor suppressor gene that encodes a protein of 199 amino acids. Expression of PP5715 in E. coli was mainly as inclusion bodies. The recombinant protein was purified by a NTA-Ni2+ affinity column, refolded by dialysis, and shown to suppress growth of two human hepatocellular carcinoma cell lines using the MTT assay or the clonogenic assays. Computer analysis and fusion expression of PP5175 and GFP showed that PP5715 may be a secretory protein. It may bind to receptors on the membrane surface and thus induce regulation of cell growth. These preliminary results suggest that protein PP5715 may be a new tumor suppressor with growth inhibition effects on hepatocellular carcinoma cells.
Wen-Zhe
Ma
M
Research Center of Biotechnology, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200233, China.
Wei
Han
H
Wen-Xin
Qin
Q
Da-Fang
Wan
W
Jian-Ren
Gu
G
Sheng-Li
Yang
Y
Yi
Gong
G
D001483:Base Sequence; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D002455:Cell Division / Q000235:genetics*; D045744:Cell Line, Tumor; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D016147:Genes, Tumor Suppressor; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D013347:Subcellular Fractions / Q000378:metabolism
D017931:DNA Primers; D011994:Recombinant Proteins