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- W1521414324 abstract "AIM: To clone and express a new gene psl12 from shark liver, purify the expression product, and study its inhibitory effects on hepatoma cells. METHOD AND RESULT: We have previously reported the discovery of Hepatocyte regenerational stimulatory factor from shark liver (sHRSF). According to the sequence of the N-terminal amino acid residues of sHRSF and using RT-PCR method, we obtained 350 bp cDNA fragment from the total RNA extracted from regenerated hepatic tissues. The cDNA has an ORF of 333 nucleotides and encodes a peptide of 111 amino acid residues. The front 7 N-terminal amino acid residues agree with those of sHRSF. We named this peptide PSL12 (peptide from shark liver with a molecular mass of about 12 kDa). The cDNA was ligated into plasmid pGEM-T-Easy and obtained recombinant plasmid pGEM-T-psl12. Based on the sequence of psl12 gene and multiple cloning sites (BamH Ⅰ and Sal Ⅰ restriction sites) of expression plasmid pET-32a, the specific primers for PCR amplifying psl12 gene were designed and synthesized. The PCR was operated using plasmid pGEM-T-psl12 as template. The expression plasmid pET-32a-psl12 was constructed by inserting the PCR product into pET-32a and was transformed into an E. coli host cell BL21. After inducing with IPTG, the fusion protein with his-tag was expressed to about 40% and was purified using metal chelation chromatography on His-Bind resin. After the fusion protein was cleaved with FXa, PSL12 was purified using Resource Q and Mono Q column chromatography. The purified PSL12 could inhibit proliferation of the hepatoma cell lines SMMC-7721 and HepG2. CONCLUSION: A new gene psl12 was obtained from shark liver. The expression product PSL12 of the gene could inhibit proliferation of the hepatoma cells cultured in vitro." @default.
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- W1521414324 date "2009-01-01" @default.
- W1521414324 modified "2023-09-26" @default.
- W1521414324 title "Cloning and Expression of a New Gene from Shark Liver and its Inhibitory Effects on Hepatoma Cells" @default.
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- W1521414324 doi "https://doi.org/10.1016/s1875-5364(09)60044-3" @default.
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